HK1181035A - 5-(biphenyl-4-yl)-3-phenyl-1,2,4-oxadiazolyl derivatives as ligands on the sphingosine 1-phosphate (s1p) receptors - Google Patents

Description

5- (biphenyl-4-yl) -3-phenyl-1, 2, 4-oxadiazolyl derivatives as ligands on sphingosine-1-phosphate (S1P) receptors

The present invention relates to chiral oxadiazoles, their use as pharmaceuticals and their use in the treatment of multiple sclerosis and other diseases.

In particular, the invention relates to compounds of formula (I) and groups thereinIs oxidized intoOxidized forms, pharmaceutically acceptable derivatives of (A),Solvates, tautomers, salts, enantiomers, diastereomers, including mixtures thereof in all ratios:

wherein

R¹Represents H, halogen, CF₃、OCF₃CN or NO₂、OH、A、OA

X¹、X²、Z¹、Z²Independently of one another, H or A, wherein X¹、X²、Z¹、Z²At least one of which is a,

X¹、X²can be reacted with C₁Together form a 3-8 membered ring,

Z¹、Z²can be reacted with C₂Together form a 4-5 membered ring,

Y¹、Y²h, A or glucuronide (glucuronide) independently of each other,

W¹、W²、W³and W⁴Independently of one another represent-CH₂-a group or a single bond.

R^aRepresents halogen or a linear or branched alkyl group having 1 to 3 carbon atoms, wherein 1 to 3H atoms may be replaced by halogen.

R^bRepresents halogen, straight-chain or branched alkyl having 1 to 3 carbon atoms, in which 1 to 3H atoms may be replaced by halogen, -OCH₃、-OCH₂CH₃Instead of this, the user can,

G¹、G²independently of one another, H, halogen, A,

a represents a linear or branched alkyl group having 1 to 6 carbon atoms, preferably 1 to 3 carbon atoms, in which 1 to 3H atoms may be substituted by halogenElement, OCH₃And the substitution of OH is carried out,

C₁、C₂is a carbon atom.

In another embodiment, C₁Or C₂Neither is chiral. In other words, X¹And X²Are identical, and Z¹And Z²Are the same.

In another embodiment, C₁And C₂Is a chiral center. X¹And X²And/or Z¹And Z²Is different.

In one embodiment, the invention provides as one diastereomer a compound of formula (I) wherein C₁Is (R) and C₂Is (R), or wherein C₁Is (S) and C₂Is (S), or a mixture thereof in all proportions.

In another embodiment, the invention provides as one diastereomer a compound of formula (I) wherein C₁Is (S) and C₂Is (R), or wherein C₁Is (R) and C₂Is (S), or a mixture thereof in all proportions.

In another embodiment, the present invention provides a compound of formula (I) as enantiomer IA, wherein C ₁Is (S) and C₂Is (R).

In another embodiment, the invention provides as enantiomer IB, a compound of formula (I) wherein C₁Is (R) and C₂Is (S).

In another embodiment, the invention provides a compound of formula (I) as enantiomer IC, wherein C₁Is (S) and C₂Is (S).

In another embodiment, the invention provides as enantiomer ID a compound of formula (I) wherein C₁Is (R) and C₂Is (R).

At C₁And C₂In the case where only one of them is a chiral centre, the invention may provide as one enantiomer a compound of formula (I) wherein at C₁Or C₂The chiral center at (A) is (S). Alternatively, the invention may provide as one enantiomer a compound of formula (I) wherein at C₁Or C₂The chiral center at (A) is (R). Wherein C is₁And C₂Compounds of formula (I) which are chiral in one aspect are also available as mixtures of enantiomers in all ratios, including racemic mixtures in which the ratio is 50/50. When the compound of formula (I) is obtained as an optically active compound, the ratio of one enantiomer to the other is higher than about 60/40, preferably higher than about 80/20, more preferably higher than about 90/10, more preferably higher than about 95/5, even more preferably higher than 98/2.

In another embodiment, enantiomer IA (wherein C) of a compound of formula (I)₁Is (S) and C₂Is (R)) with enantiomer IB of a compound of formula (I) (wherein C₁Is (R) and C₂Is (S)) in an arbitrary ratio. When the compound of formula (I) is obtained as racemate, the two enantiomers IA and IB are mixed in equal amounts. When the compound of formula (I) is obtained as optically active compound, the ratio between enantiomer IA and enantiomer IB is higher than about 60/40, preferably higher than about 80/20, more preferably higher than about 90/10, more preferably higher than about 95/5, even more preferably higher than 98/2.

In another embodiment, the enantiomer IC of the compound of formula (I) (wherein C₁Is (S) and C₂Is (S)) with the enantiomer ID of the compound of formula (I) wherein C₁Is (R) and C₂Is (R)) in an arbitrary ratio. When the compound of formula (I) is obtained as an optically active compound, the ratio between enantiomer IC and enantiomer ID is higher than about 60/40, preferably higher than about 80/20, more preferably higher than about 90/10, more preferably higher than about 95/5, even more preferably higher than 98/2.

In a preferred embodiment, the ratio of one enantiomer to the other is higher than 98/2.

In one embodiment, the compound of formula (I) is an enantiomer, wherein:

a) X¹、X²and Z¹Is H and Z²Is a group A of a group A,

b) X¹、X²and Z²Is H and Z¹Is a group A of a group A,

c) Z¹、Z²and X²Is H and X¹Is A, or

d) Z¹、Z²And X¹Is H and X²Is a group A of a group A,

either alone or in combination with one or more others in all proportions.

In another embodiment, the compound of formula (I) is a diastereomer, wherein:

a) X¹and Z¹Is H, X²And Z²Is A and/or X²And Z²Is H, X¹And Z¹Is A, or

b) X¹And Z²Is H, X²And Z¹Is A and/or X²And Z¹Is H, X¹And Z²Is a group A of a group A,

either alone or in combination in all proportions.

An enantiomerically enriched mixture (enantiomerically enriched mixture) represents a compound of formula (I) or a related formula having the following enantiomeric excess as measured by methods well known to those skilled in the art: 10% or more, preferably 50% or more, and more preferably more than 95%. Most preferably, an enantiomerically enriched mixture represents a compound of formula (I) or related formulae having an enantiomeric excess of more than 98%.

In another embodiment, the invention provides wherein W¹、W²、W³And W⁴At least 1 or 2 of which are single bonds.

In another embodiment, W¹、W²、W³And W⁴All represent single bonds.

In another embodiment, the present invention provides wherein Z is¹And Z²Can be reacted with C₂Compounds of formula (I) which together form a 3-8 membered ring.

In another embodiment, the present invention provides a compound of formula (Ia)

Wherein G is¹、G²、R^a、R^b、R¹、X¹、X²、Z¹、Z²And Y¹As defined above.

In another embodiment, the present invention provides a compound of formula (Ib)

Wherein G is¹、R^a、R^b、R¹、X¹、X²、Z¹、Z²And Y¹As defined above.

In one embodiment, the present invention provides a compound of formula (I), (Ia) or (Ib), wherein R^bIs CF₃Or CH₂OCH₃And R is^aIs CF₃Or CH₃Preferably, R^bIs CF₃And R is^aIs CH₃。

In another embodiment, the present invention provides a compound of formula (I), (Ia) or (Ib), wherein R^bis-CF₃or-CH₂OCH₃And R is^ais-CF₃or-CH₃Preferably, R^bis-CF₃And R is^ais-CH₃。

In another embodiment, the present invention provides a compound of formula (I), (Ia) or (Ib), wherein R^bis-CF₃or-CH₂OCH₃And R is^ais-CF₃or-CH₃Preferably, R^bis-CF₃And R is^ais-CH₃。

In another embodiment, the present invention provides a compound of formula (I) or (Ia), wherein R^bis-CF₃or-CH₂OCH₃And R is^ais-CF₃or-CH₃Preferably, R^bis-CF ₃And R is^ais-CH₃And Y is²Is H.

In another embodiment, the present invention provides a compound of formula (I), (Ia) or (Ib), wherein R^aAnd R^bOne is-CF₃。

In another embodiment, the present invention provides compounds of formula (Ic) and pharmaceutically acceptable derivatives, solvates, tautomers, salts and stereoisomers thereof, including mixtures thereof in all ratios

Wherein R is¹、R^a、G¹、G²、W¹、W²、W³、W⁴、X¹、X²、Z¹、Z²、Y¹And Y²As defined aboveAnd (5) defining.

In another preferred embodiment, the present invention provides a compound of formula (Id)

Wherein R is¹、R^b、G¹、G²、W¹、W²、W³、W⁴、X¹、X²、Z¹、Z²、Y¹And Y²As defined above.

In another embodiment, the present invention provides a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R^bis-CH₃、-CH₂CH₃F, Br, Cl or-CF₃Preferably F, -CH₃or-CF₃，

G₁、G₂Independently of one another, H, halogen or-CH₃Preferably H.

And wherein R is^b、W、R¹、W¹、W²、W³、W⁴、X¹、X²、Z¹、Z²、Y¹And Y²As defined above.

In a more preferred embodiment, the present invention provides a compound of formula (Ie):

wherein R is^a、G¹、G²、X¹、X²、Z¹、Z²、Y¹As defined above.

In a more preferred embodiment, the present invention provides a compound of formula (If):

wherein R is^a、G¹、G²、X¹、X²、Z¹、Z²、Y¹As defined above.

In another preferred embodiment, the present invention provides a compound of formula (Ig):

Wherein R is¹、R^b、G¹、G²、X¹、X²、Z¹、Z²、Y¹As defined above.

In another preferred embodiment, the present invention provides a compound of formula (Ih):

wherein R is¹、R^b、G¹、G²、X¹、X²、Z¹、Z²、Y¹As defined above.

In another embodiment, the present invention provides a compound of formula (I), (Ia), (Ib), (Id), (Ig) or (Ih) wherein R is^bis-CH₂OCH₃。

Wherein Y is¹And/or Y²The compound of formula (I) which is glucuronide may be, for example, a compound of formula (Ik):

wherein G is¹、G²、R^a、R^b、R¹、W¹、W²、W³、W⁴、X¹、X²、Y¹、Z¹And Z²As defined above.

The oxidised form of the compound of formula (I) may be, for example, a compound of formula (IO):

wherein G is¹、G²、R^a、R^b、R¹、W¹、W²、W³、W⁴、X¹、X²、Y¹、Y²、Z¹And Z²As defined above.

The compounds of formula (I) and related formulae preferably bind to the receptor for sphingosine 1-phosphate (S1P). S1P is a bioactive sphingolipid metabolite secreted by hematopoietic cells and stored and released from activated platelets. It acts as an agonist on the family of G protein-coupled receptors (GPCRs). 5 sphingosine 1-phosphate receptors have been identified (S1P)₁、S1P₂、S1P₃、S1P₄And S1P₅Also known as endothelial differentiation genes (endoglin differentiation genes), which are Edg1, Edg5, Edg3, Edg6 and Edg8, respectively, have a broad cellular and tissue distribution and are well conserved in human and rodent species.

S1P is involved in many cellular functions such as survival, proliferation, and immune response. The compounds of the invention preferably have the general formula S1P₁The effects of/Edg 1 receptor agonists and by modulating leukocyte trafficking, sequestering lymphocytes in secondary lymphoid tissues, and interfering with the cell-cell interactions required for an effective immune responseHas immunosuppressive activity. The invention also relates to pharmaceutical compositions containing such compounds and methods of treatment or prevention.

FTY720 or Fingolimod (a non-selective S1P)₁Agonist) produces immunosuppressive activity and exhibits therapeutic effects in the treatment of relapsing-remitting multiple sclerosis. Numerous publications have been published using this compound: cyster JG Annu Rev Immunol 23:127-59, 2005, Rosen H Nat Rev Immunol 5: 560-.

Immunosuppressants can further be used in a variety of autoimmune and chronic inflammatory diseases, including multiple sclerosis, systemic lupus erythematosus, chronic rheumatoid arthritis, type I diabetes, inflammatory bowel disease, biliary cirrhosis, uveitis, and other conditions such as crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis, autoimmune myositis, wegener's granulomatosis, ichthyosis, graves ophthalmopathy, atopic dermatitis, and asthma. They are also used as part of chemotherapeutic regimens for the treatment of cancer, lymphoma and leukemia.

Patent application WO2006/131336 describes oxadiazole derivatives containing a biphenyl ring. Other oxadiazole derivatives containing a phenyl group substituted by a cycloalkyl group are known from Bioorg med. chem. 16 (2006) 3679-3683.

Other oxadiazole derivatives are described in patent application EP 07117921.2.

It has been found that the compounds of the invention are selective S1P with improved drug-like and/or other properties₁An agonist.

The invention uses compounds of formula (I) and pharmaceutically acceptable derivatives, salts, tautomers, solvates and stereoisomers thereof, including mixtures thereof in all ratios, for the preparation of medicaments for use in therapy and/or preventing S1P therein₁Diseases in which inhibition, activation, modulation and/or modulation of receptor signaling plays a role.

Thus, the present invention preferably comprises compounds that: they are S1P₁Agonists of the/Edg 1 receptor, in particular having advantages over the S1P₃Selective agonists of the/Edg 3 receptor. S1P₁the/Edg 1 receptor selective agonists have advantages over existing therapies and extend the therapeutic window of lymphocyte spacers, thus allowing better tolerability at higher doses and thus improved efficacy.

The invention further relates to the preparation of medicaments for improving vascular function, alone or in combination with other active compounds or therapies.

The invention further relates to a set or kit consisting of the following individual packages:

(a) an effective amount of a compound according to formula (I) or related formulae and/or pharmaceutically acceptable derivatives, tautomers, salts, solvates and stereoisomers thereof, including mixtures thereof in all ratios,

and

(b) an effective amount of other pharmaceutically active ingredients.

The following abbreviations respectively represent the following definitions:

aq (aqueous), h (hours), g (g), L (L), mg (mg), MHz (megahertz), μ M (micromolar), min. (minutes), mM (millimeters), mmol (millimolar), mM (millimolar), m.p. (melting point), eq (equivalent), mL (mL), μ L (microliter), ACN (acetonitrile), BINAP (2,2 '-bis (diphenylphosphino) -1, 1' -binaphthalene, BOC (tert-butoxy-carbonyl), CBZ (benzyloxycarbonyl), CDCl3 (deuterated chloroform), CD (r-butyl ether), hi (r-₃OD (deuterated methanol), CH₃CN (acetonitrile), c-hex (cyclohexane), DCC (dicyclohexylcarbodiimide), DCM (dichloromethane), dppf (1,1' -bis (diphenylphosphino) ferrocene), DIC (diisopropylcarbodiimide), DIEA (diisopropylethyl-amine), DMF (dimethyl formamide) Formamide), DMSO (dimethyl sulfoxide), DMSO-d₆(deuterated dimethyl sulfoxide), EDC (1- (3-dimethyl-amino-propyl) -3-ethylcarbodiimide), ESI (electrospray ionization), EtOAc (ethyl acetate), Et₂O (diethyl ether), EtOH (ethanol), FMOC (fluorenylmethyloxycarbonyl), HATU (dimethylamino- ([1,2, 3)]Triazolo [4,5-b]Pyridin-3-yloxy) -methylene]-dimethyl-ammonium hexafluorophosphate), HPLC (high performance liquid chromatography), i-PrOH (2-propanol), K₂CO₃(Potassium carbonate), LC (liquid chromatography), MD Autoprep (mass-directed Autoprep), MeOH (methanol), MgSO₄(magnesium sulfate), MS (Mass Spectrometry), MTBE (methyl Tert-butyl Ether), mtr. (4-methoxy-2, 3, 6-trimethylbenzenesulfonyl), MW (microwave), NBS (N-bromosuccinimide), NaHCO₃(sodium bicarbonate), NaBH₄(sodium borohydride), NMM (N-methylmorpholine), NMR (nuclear magnetic resonance), POA (phenoxy acetate), Py (pyridine), PyBOP (benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate), RT (room temperature), RT (retention time), SFC (supercritical fluid chromatography), SPE (solid phase extraction), T3P (propylphosphonic anhydride), TBAF (tetra-N-butylammonium fluoride), TBTU (2- (1-H-benzotriazol-1-yl) -1,1,3, 3-tetramethyluronium tetrafluoroborate), TEA (triethylamine), TFA (trifluoroacetic acid), THF (tetrahydrofuran), TLC (thin layer chromatography), UV (ultraviolet).

In general, the chiral oxadiazole compounds according to formula (I) and related formulae of the present invention can be prepared from readily available starting materials. If such starting materials are not commercially available, they can be prepared by standard synthetic techniques. In general, the synthetic route for any individual compound of formula (I) and related formulae will depend on the particular substituents per molecule, such factors being known to those of ordinary skill in the art. The following general methods and procedures, described below in the examples, may be used to prepare compounds of formula (I) and related formulae. The reaction conditions, such as temperature, solvents or co-reagents (co-reagents), described in the following schemes are given as examples only and are not limiting. It is to be understood that where typical or preferred experimental conditions (i.e., reaction temperature, time, moles of reagents, solvents, etc.) are given, other experimental conditions may also be used, unless otherwise specified. Optimal reaction conditions may vary with the particular reactants or solvents used, but such conditions may be determined by one skilled in the art using routine optimization procedures. For all protection and deprotection methods see Philip J. Kocienski, "Protecting Groups", Georg Thieme Verlag Stuttgart, New York, 1994, and Theodora W. Greene and Peter G.M. Wuts, "Protective Groups in Organic Synthesis", Wiley Interscience, 3 rd edition 1999.

According to R¹、R^a、R^b、G¹、G²、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹And Z²Different synthetic strategies can be chosen for the synthesis of the compounds of formula (I). In the methods illustrated in the schemes below, R¹、R^a、R^b、G¹、G²、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹And Z²As defined in the description above, unless otherwise mentioned.

Typically, by hydrolysis of an ester derivative of formula (I) (wherein Y is²As defined above, and more preferably Y²Is methyl or tert-butyl) can be used to prepare compounds of formula (I') (wherein R is¹、R^a、R^b、G¹、G²、W¹-W⁴、X¹、X²、Y¹、Z¹And Z²As defined above, and Y²Is H), using conditions well known to those skilled in the art, such as metal hydroxide (e.g. lithium hydroxide, sodium hydroxide or potassium hydroxide), in a suitable solvent (such as THF, methanol, ethanol or water or mixtures thereof), or using an acid (e.g. HCl or TFA), in a suitable solvent (such as dioxane, DCM), at a temperature of about 20 ℃ to about 50 ℃, preferably at room temperature, for several hours, e.g. 1 hour to 24 hours (scheme)1)。

Scheme 1

When the ester derivatives of formula (I) are obtained as a mixture of enantiomers, they may be separated by a chiral HPLC column, such as, but not limited to, the methods described below in the examples.

The process for the preparation of ester derivatives of formula (I) selected from the following is more particularly described in the examples:

2- [ (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) (methyl) amino ] butanoic acid tert-butyl ester,

methyl N- [ (1R) -1- (4- {5- [2' - (difluoromethyl) -2-methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

methyl N- [ (1S) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

methyl N- [ (1R) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

n- [ (1S) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

methyl N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

n- [ (1R) -1- (4- {5- [2' - (difluoromethyl) -2-methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

n- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

n- [ (1R) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

Ethyl (2S) -2- { [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] amino } butanoate,

ethyl (2R) -2- { [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] amino } butanoate,

n- (3-fluoro-5- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) leucine tert-butyl ester,

n- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -N-methylvaline tert-butyl ester,

N-methyl-N- [1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

n- [1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -N-methylglycine tert-butyl ester,

tert-butyl N- (2-fluoro-4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine,

methyl N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

methyl N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine,

Methyl N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine,

methyl N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

n- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester

N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

n- [1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

n- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine methyl ester

Methyl N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alaninate,

methyl N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine,

N-methyl-N- [1- (3- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] alanine ester,

N-methyl-N- [1- (3- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] alanine ester,

N-methyl-N- [1- (3- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

tert-butyl N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -N, 2-dimethylalaninate,

n- [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

N-methyl-N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

N-methyl-N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

N-methyl-N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

N-methyl-N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

methyl N- [1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -N-methylalanine,

n- [1- (3- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

Methyl N-methyl-N- [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] alanine,

N-methyl-N- [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

2- [ (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) amino ] butanoic acid tert-butyl ester,

n- [1- (3- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

n- (2-fluoro-4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -N, 2-dimethylalanine tert-butyl ester,

n- (2-fluoro-4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -N-methylalanine tert-butyl ester,

o- (tert-butyl) -N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -N-methylserine tert-butyl ester,

tert-butyl N- (3-fluoro-5- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine,

tert-butyl N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) valine,

Tert-butyl N- (2-fluoro-4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine,

tert-butyl N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine,

n- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) -N-methylalanine tert-butyl ester,

n- [ 2-methoxy-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -N-methylglycine tert-butyl ester,

n- [ 2-hydroxy-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] glycine tert-butyl ester,

3-methyl-2- (4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenethylamino) butanoic acid- (2S) -tert-butyl ester,

n- [ (1R) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -N-methylglycine tert-butyl ester,

n- [ (1S) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -N-methylglycine tert-butyl ester,

tert-butyl N- [ 2-hydroxy-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine,

Tert-butyl N- [2- { [ tert-butyl (dimethyl) silyl ] oxy } -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine.

According to a preferred synthetic route, the compound of formula (I) wherein R is R can be obtained in a two-step process as shown in scheme 2¹、R^a、R^b、G¹、G²、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹And Z²As defined above. The first step consists of: reacting a carboxylic acid of formula (II) (wherein R^a、R^b、G¹And G²As defined above) with an ammoxim of formula (III) (wherein R is¹、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹And Z²As defined above). A general scheme for such coupling is given below in the examples, using conditions and methods well known to those skilled in the art. Standard coupling agents (such as HATU, EDC or isobutyl chloroformate) may be used in the presence or absence of a base (such as DIEA, TEA or NMM) in a suitable solvent (such as DMF, ACN, THF or iPrOAc) at temperatures from about 0 ℃ to room temperature (preferably at 0 ℃) for periods of 30 minutes to several hours. Alternatively, a carboxylic acid derivative (e.g., an acid chloride) can be coupled with an ammoxim (III) for several hours using conditions and methods well known to those skilled in the art in the presence of a base (such as pyridine or DIEA) in a suitable solvent (such as toluene, DCM, THF or DMF) at a temperature of about 0 ℃ to room temperature (preferably at room temperature). The second step consists of: cyclizing and dehydrating the O-substituted ammoxim (IV) to form the oxadiazole (I). Conditions are given in the examples below, where the oxadiazole is prepared using methods well known to those skilled in the art, such as pyrolysis (thermolysis) in the presence or absence of a base (such as DIEA, TEA or NMM) in a suitable solvent or solvent mixture (such as toluene, pyridine, ACN, THF, DMF or iPrOAc) at a temperature of 80 ℃ to about 120 ℃, typically 90 ℃, for 12-72 hours (preferably 15 hours).

Scheme 2

Alternatively, the alcohol derivative of formula (V) may be converted to the corresponding amine derivative of formula (I), as shown in scheme 3. The alcohol function of the compound of formula (V) may be first converted to a leaving group, such as a chloride or sulfonate (sulfonate), using conditions well known to those skilled in the art. For example, the alcohol derivative of formula (V) may be reacted with methanesulfonyl chloride in the presence of a base such as, but not limited to, a tertiary amine (e.g., TEA or DIEA) in a suitable solvent such as DCM at a temperature of about 20 ℃ to about 50 ℃ (preferably at room temperature) for several hours. The resulting compound is then reacted with a suitable compound of formula HN (Y)¹)W³C²(Z¹)(Z²)W⁴COOY²To give the compound of formula (I). Alternatively, conditions well known to those skilled in the art are used, such as, but not limited to, Swern oxidation conditions, or MnO is used₂As an oxidizing agent for benzyl alcohol (benzyl alcohols), the alcohol derivatives of formula (V) can be oxidized to the corresponding aldehydes. With a suitable formula HN (Y) in the presence of a reducing agent such as, but not limited to, sodium cyanoborohydride¹)W³C²(Z¹)(Z²)W⁴COOY²The resulting aldehyde is reductively aminated with an amine of (a) to give a compound of formula (I).

Scheme 3

Using a suitable ammoxim of formula (III') (wherein R ¹、W¹、W²、X¹And X²As defined above) may be derived from carboxylic acids of formula (II) (wherein R is^a、R^b、G¹And G²As defined above) to prepare the alcohol derivative of formula (V).

When the alcohol derivatives of formula (V) are obtained as a mixture of enantiomers, they may be separated by a chiral HPLC column, such as, but not limited to, the methods described below in the examples.

The process for the preparation of alcohol derivatives of formula (V) selected from the following is more particularly described in the examples:

1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethanol,

(4- (5- (2- (methoxymethyl) -2' -methylbiphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) methanol,

(4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) methanol,

(3- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) methanol,

(4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) ethanol,

(3- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) ethanol.

Alternatively, the amine derivative of formula (V') can be converted to the corresponding amine derivative of formula (I), as shown in scheme 3 a. The amine of formula (V') may be reacted with a suitable amine of formula LG- (Y) ¹)W³C²(Z¹)(Z²)W⁴COOY²Wherein LG is a leaving group. A preferred leaving group is a halogen, such as but not limited to Br. Such a reaction will yield a compound of formula (I).

Scheme 3a

Use of a suitable ammoxim of formula (III '') (wherein R¹、W¹、W²、X¹And X²As defined above, and PG is a protecting group, such as but not limited to boc), can be derived from a carboxylic acid of formula (II) (wherein R is^a、R^b、G¹And G²As defined above) to prepare the amine derivative of formula (V') as shown in scheme 3 b. The resulting amine derivative of formula (V ") can be converted to an amine derivative of formula (V') using a deprotection step.

Scheme 3b

When the derivatives of formula (V') or (V ") are obtained as a mixture of enantiomers, they may be separated by a chiral HPLC column, such as, but not limited to, the methods described below in the examples.

The process for the preparation of amine derivatives of formula (V') selected from the following is more particularly described in the examples:

(1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethanamine

N-methyl-1- (3- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) methanamine.

The process for the preparation of the derivatives of formula (V ") selected from the following is more particularly described in the examples:

[ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] carbamic acid tert-butyl ester

Methyl (3- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) carbamic acid tert-butyl ester.

The process for the preparation of an ammoxim of formula (III ") selected from the following is more particularly described in the examples:

((1R) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) carbamic acid tert-butyl ester

{3- [ amino (hydroxyimino) methyl ] benzyl } methylcarbamic acid tert-butyl ester.

Compounds of formula (II) (wherein R is R) may be prepared by standard synthetic techniques as described hereinafter in the examples using conditions and procedures well known to those skilled in the art (scheme 4)^a、R^b、G¹And G²As defined above). In the first synthetic route, the compound of formula (II) (wherein R is^a、R^b、G¹And G²As defined above). More particularly, by alkyl benzoates (VII) (wherein LG₁It may be preferred that they are obtainable by a Suzuki-Miyura coupling reaction between Br, I or a sulfonate such as triflate (triflate) and a boronic acid or ester of formula (VIII) (Miyaura, N.; Suzuki, A. chem. Rev. 1995, 95, 2457; Takahiro I. and Toshiaki M., Tetrahedron Lett. 2005, 46, 3573-one 3577). In a typical procedure, in a suitable solvent (such as a mixture of toluene and water), in the presence of a base (such as K) ₂CO₃) And a catalytic amount of a palladium catalyst (such as Pd (PPh)₃)₄) In the presence of, and optionally adding, a phosphine ligand (such as PPh)₃) The alkyl benzoate (VII) and the boric acid (VIII) are heated. The resulting ester (VI) can be hydrolyzed using a metal hydroxide (such as NaOH) in a suitable solvent (such as MeOH, EtOH, water or mixtures thereof) at a temperature of about 20 ℃ to 60 ℃ (preferably at room temperature) for several hours.

Scheme 4

In a second synthesis route, the compound of formula (II) (in which R is R) can be obtained by coupling reaction followed by hydrolysis of the nitrile (IX) obtained^a、R^b、G¹And G²As defined above). The resulting benzonitrile of formula (IX) can be hydrolyzed to the corresponding carboxylic acid (II) by treatment with an aqueous solution of a metal hydroxide, such as NaOH, in a suitable solvent, such as MeOH or EtOH, at temperatures ranging from room temperature to reflux, preferably at reflux, for several hours, for example, 1-24 hours.

In a third synthetic route, according to scheme 4, compounds of formula (II) (wherein R is R) can be prepared from aryl bromides of formula (XI) in a two-step process^a、R^b、G¹And G²As defined above). The first step is: in a suitable solvent (such as Et)₂O), at low temperature (preferably at-78 deg.C) with lithiated alkyl (such as nBuLi ortBuLi) for halogen-metal exchange. The second step is: by adding CO as electrophile₂The organolithiated derivative is quenched (as a gas or in the solid state).

When the compounds of formula (II) are obtained as a mixture of enantiomers, they may be separated by a chiral HPLC column, such as, but not limited to, the methods described below in the examples.

The process for the preparation of a compound of formula (II) selected from the following is more particularly described in the examples:

2' -methyl-2- (trifluoromethyl) biphenyl-4-carboxylic acid,

2' - (difluoromethyl) -2-methylbiphenyl-4-carboxylic acid,

2- (methoxymethyl) -2' -methylbiphenyl-4-carboxylic acid,

2-methyl-2' - (trifluoromethyl) biphenyl-4-carboxylic acid.

A compound of formula (III) (wherein R¹、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹And Z²As defined above) are commercially available or can be prepared according to scheme 5 as follows: in the presence or absence of a base (such as TEA) in the chamberTo a temperature of about 80 c (preferably at room temperature), an aqueous solution of hydroxylamine or hydroxylamine hydrochloride is added to a solution of the corresponding substituted benzonitrile of formula (XII) in a suitable solvent such as EtOH for several hours.

Scheme 5

When the compounds of formula (III) are obtained as a mixture of enantiomers, they may be separated by a chiral HPLC column, such as, but not limited to, the methods described below in the examples.

The process for the preparation of the compounds of formula (III) selected from the following is more particularly described in the examples:

2- [ {4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } (methyl) amino ] butyric acid tert-butyl ester,

n- ((1R) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -D-alanine methyl ester,

n- [ (1R) -1- (4-cyanophenyl) ethyl ] -L-alanine methyl ester,

n- ((1S) -1- {4- [ (hydroxyamino) (imino) methyl ] phenyl } ethyl) glycine tert-butyl ester,

n- ((1R) -1- {4- [ (hydroxyamino) (imino) methyl ] phenyl } ethyl) glycine tert-butyl ester,

n- ((1S) -1- {4- [ (hydroxyamino) (imino) methyl ] phenyl } ethyl) -D-alanine methyl ester,

n- ((1S) -1- {4- [ (hydroxyamino) (imino) methyl ] phenyl } ethyl) -L-alanine methyl ester,

(2S) -2- [ ((1R) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) amino ] butanoic acid ethyl ester,

ethyl (2R) -2- [ ((1R) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) amino ] butanoate,

ethyl 2- ({3- [ amino (hydroxyimino) methyl ] -5-fluorobenzyl } amino) -4-methylpentanoate,

n- {4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } -N-methylvaline tert-butyl ester,

n- (1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylglycine tert-butyl ester,

tert-butyl 2- ({4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } amino) -2-methylpropionate,

(1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) carbamic acid tert-butyl ester,

n- (1- {3- [ (hydroxyamino) (imino) methyl ] phenyl } ethyl) -N-methylalanine methyl ester,

n- (1- {3- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylglycine (methyl-tert-butyl) ester,

2- [ {4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } (methyl) amino ] -2-methylpropanoic acid tert-butyl ester,

n- (1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) glycine tert-butyl ester,

n- ((1S) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylglycine tert-butyl ester,

n- ((1R) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylglycine tert-butyl ester,

n- (1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylalanine methyl ester,

n- (1- {3- [ amino (hydroxyimino) methyl ] phenyl } ethyl) glycine tert-butyl ester,

tert-butyl 2- ({4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } amino) butanoate,

2- [ {4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } (methyl) amino ] -2-methylpropanoic acid tert-butyl ester,

tert-butyl 2- [ {4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } (methyl) amino ] propionate,

n- {4- [ (amino (hydroxyimino) methyl ] -2-fluorobenzyl } -O- (tert-butyl) -N-methylserinic acid tert-butyl ester,

tert-butyl 2- ({3- [ amino (hydroxyimino) methyl ] -5-fluorobenzyl } amino) -2-methylpropionate,

Tert-butyl 2- ({4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } amino) -3-methylbutyrate,

tert-butyl 2- ({4- [ amino (hydroxyimino) methyl ] -2-fluorobenzyl } amino) -2-methylpropionate,

tert-butyl 2- ((1- (4- (N' -hydroxycarbamimidoyl) phenyl) -2-methoxyethyl) (methyl) amino) acetate,

tert-butyl 2- (2- (tert-butyldimethylsilyloxy) -1- (4- (N' -hydroxycarbamimidoyl) phenyl) ethylamino) acetate,

n' -hydroxy-4- (hydroxymethyl) benzamidine,

n' -hydroxy-3- (hydroxymethyl) benzamidine,

n' -hydroxy-4- (hydroxyethyl) benzamidine,

n' -hydroxy-3- (hydroxyethyl) benzamidine,

tert-butyl N- (1- {4- [ (Z) -amino (hydroxyimino) methyl ] phenyl } -2- { [ tert-butyl (dimethyl) silyl ] oxy } ethyl) -D-alanine.

A compound of formula (XII) (wherein R¹、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹And Z²As defined above) are commercially available or may be prepared from alternative compounds of formula (XII) as follows: suitable interconversion procedures are used, such as those described in the examples below, or conventional interconversion procedures well known to those skilled in the art. Alternatively, but not limited thereto, the compound of formula (XII) may be prepared from an aldehyde or ketone of formula (XIIa) as follows: with a suitable formula HN (Y) in the presence of a reducing agent such as, but not limited to, sodium cyanoborohydride ¹)W³C²(Z¹)(Z²)W⁴COOY²The amine of (a) is subjected to reductive amination. Alternatively, an aldehyde or ketone of formula (XIIb) may be converted to a chiral imine (XIIc) by reaction with a chiral auxiliary, such as but not limited to a tert-butane sulfenamide group, in the presence of titanium ethoxide (Ellman j.a. et al,Acc. Chem. Res. 2002, 35, 984-995). It can be further converted to the sulfenamide (XIId) by reduction or nucleophilic addition using conditions known to those skilled in the art. The sulfinyl group is then removed by acid treatment (such as HCl in a protic solvent) to give the desired chiral amine (XIIe) as the hydrochloride salt. The absolute configuration of the amine (XIIe) depends on the configuration of the chiral auxiliary and the conditions used for the reduction or nucleophilic addition. For example, imine (XIIc) can be reduced diastereoselectively with a reducing agent, such as, but not limited to, sodium borohydride or lithium tri-sec-butylborohydride (L-selectride), to give chiral amine (XIIe) after removal of the chiral auxiliary agent (Ellman j.a. et al,J. Org. Chem. 2007, 72,626-629). The chiral amine (XIIe) may be further converted to a compound of formula (XII) by reductive alkylation with a suitable aldehyde or ketone, or by direct alkylation with a suitable electrophile such as an alkyl bromide or sulfonate derivative: .

Scheme 6

When the compounds of formula (XII) are obtained as a mixture of enantiomers, they may be separated by a chiral HPLC column, such as, but not limited to, the methods described below in the examples.

The process for the preparation of a compound of formula (XII) selected from the following is more particularly described in the examples:

tert-butyl 2- [ (2-fluoro-4-isocyanobenzyl) (methyl) amino ] butyrate,

n- [ (1R) -1- (4-cyanophenyl) ethyl ] -D-alanine methyl ester and N- [ (1R) -1- (4-cyanophenyl) ethyl ] -L-alanine methyl ester,

n- [ (1S) -1- (4-cyanophenyl) ethyl ] glycine tert-butyl ester,

n- [ (1R) -1- (4-cyanophenyl) ethyl ] glycine tert-butyl ester,

n- [ (1S) -1- (4-cyanophenyl) ethyl ] -D-alanine methyl ester and N- [ (1S) -1- (4-cyanophenyl) ethyl ] -L-alanine methyl ester,

ethyl (2S) -2- { [ (1R) -1- (4-cyanophenyl) ethyl ] amino } butanoate and ethyl (2R) -2- { [ (1R) -1- (4-cyanophenyl) ethyl ] amino } butanoate,

2- [ (3-cyano-5-fluorobenzyl) amino ] -4-methylpentanoic acid tert-butyl ester,

tert-butyl 2- [ (4-cyano-2-fluorobenzyl) (methyl) amino ] -3-methylbutyrate,

n- [1- (4-cyanophenyl) ethyl ] -N-methylglycine tert-butyl ester,

2- [ (4-cyano-2-fluorobenzyl) amino ] -2-methylpropanoic acid tert-butyl ester,

[1- (4-cyanophenyl) ethyl ] carbamic acid tert-butyl ester,

n- [1- (3-cyanophenyl) ethyl ] -N-methylalanine methyl ester,

n- [1- (3-cyanophenyl) ethyl ] -N-methylalanine methyl ester,

2- [ (2-fluoro-4-isocyanobenzyl) (methyl) amino ] -2-methylpropionic acid tert-butyl ester,

n- [1- (4-cyanophenyl) ethyl ] glycine tert-butyl ester,

n- [1- (4-cyanophenyl) ethyl ] -N-methylglycine tert-butyl ester,

n- [1- (4-cyanophenyl) ethyl ] -N-methylalanine methyl ester,

n- [1- (3-cyanophenyl) ethyl ] glycine tert-butyl ester,

2- [ (4-cyano-2-fluorobenzyl) amino ] butanoic acid tert-butyl ester,

2- [ (2-fluoro-4-isocyanobenzyl) (methyl) amino ] -2-methylpropionic acid tert-butyl ester,

tert-butyl 2- [ (4-cyano-2-fluorobenzyl) (methyl) amino ] propionate,

3-tert-butyl-2- [ (4-cyano-2-fluorobenzyl) (methyl) amino ] propanoate,

2- [ (3-cyano-5-fluorobenzyl) amino ] -2-methylpropanoic acid tert-butyl ester,

2- [ (4-cyano-2-fluorobenzyl) amino ] -3-methylbutyric acid-1-tert-butyl ester,

2- [ (4-cyano-2-fluorobenzyl) amino ] -2-methylpropanoic acid tert-butyl ester,

tert-butyl 2- ((1- (4-cyanophenyl) -2-methoxyethyl) (methyl) amino) acetate,

tert-butyl 2- (2- (tert-butyldimethylsilyloxy) -1- (4-cyanophenyl) ethylamino) acetate.

The compounds of formula (I) can be converted to their oxidized forms of formula (IO) using an oxidizing agent such as 3-chloroperoxybenzoic acid, as shown in scheme 7.

Scheme 7

The process for the preparation of compounds of formula (IO) selected from the following is more particularly described in the examples:

1- [ methyl (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) nitroso-methyl ] cyclopentanecarboxylic acid

2- [ (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } benzyl) (methyl) nitroso-thienyl ] propionic acid

{ methyl [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] nitroso-methyl } acetic acid.

In one embodiment, the present invention provides a process for the preparation of compounds of formula (I) and salts thereof,

reacting a compound of formula A in the presence of a suitable base (such as an amine like TEA, DIEA or NMM), or if T is OH, in the presence of a suitable condensing agent (such as EDC)

Wherein G is₁、G₂、R^aAnd R^bHas the meaning given above and T is OH or a leaving group such as Cl, Br, I, imidazolyl, pentafluorophenoxy or the reaction product of isobutyl chloroformate with formula A (wherein T is OH),

With compounds of the formula B

Wherein R is¹、W₁-W₄、C₁、C₂、X₁、X₂、Y₁、Y₂、Z₁And Z₂Having the meaning given above, it is preferred that,

and cyclizing the resulting product and optionally converting the base or acid of formula (I) into one of its salts.

The compounds of formulae (II) to (XII) may be obtained from commercial sources, or they may be prepared from known compounds as follows: operations such as those described in the examples below, or conventional operations well known to those skilled in the art, are used.

Using appropriate interconversion operations, such as those set forth hereinafterCompounds of formulae (II) to (XII) (wherein R is¹、R^a、R^b、G¹、G²、W¹-W⁴、X¹、X²、Y¹、Y²、Z¹、Z²And LG₁As defined above) to alternative compounds of formulae (II) to (XII), respectively.

If the above-mentioned general set of synthetic methods is not suitable for obtaining the compounds according to formula (I) and/or the intermediates necessary for the synthesis of the compounds of formula (I), suitable preparation methods known to the person skilled in the art should be used.

The compounds of the invention have been named according to the standard used in the program "ACD/Name Batch" (version 7.00) from Advanced Chemistry Development inc. Product version: 7.10, construction: 9/15/2003, and the program AUTONOM GOLD v 1.0.1.1.

Formula (I) and related formulae also include the optically active forms (stereoisomers), enantiomers, racemates, diastereomers and hydrates and solvates of these compounds.

Stereochemistry (S) or (R) is determined using standard rules of nomenclature well known to those skilled in the art.

According to another general method, the compounds of formula (I) and of any subformula (subformula) can be converted into alternative compounds of formula (I) and of any subformula, using suitable interconversion techniques well known to the person skilled in the art.

In general, the synthetic route of any individual compound of formula (I) will depend on: easy accessibility of specific substituents and necessary intermediates per molecule; also, such factors are known to those of ordinary skill in the art. For all protection and deprotection methods see Philip J. Kocienski,') "Protecting Groups", Georg Thieme Verlag Stuttgart, New York, 1994 and Theodora W, Greene and Peter G.M. Wuts "Protective Groups in Organic Synthesis", Wiley Interscience, 3 rd edition, 1999.

The compound of the invention bound to the solvent molecule can be isolated by crystallization by evaporation of an appropriate solvent. Pharmaceutically acceptable acid addition salts of compounds of formula (I) containing a basic centre may be prepared in a conventional manner. For example, a solution of the free base may be treated with a suitable acid (neat or in a suitable solution) and the resulting salt isolated by filtration or by evaporation of the reaction solvent under vacuum. In a similar manner, pharmaceutically acceptable base addition salts can be obtained by treating a solution of a compound of formula (I) containing an acidic centre with a suitable base. Both types of salts can be formed or interconverted using ion exchange resin techniques.

Depending on the conditions used, the reaction time is generally between a few minutes and 14 days, and the reaction temperature is between about-30 ℃ and 140 ℃, usually between-10 ℃ and 90 ℃, in particular between about 0 ℃ and about 70 ℃.

Furthermore, the compounds of formula (I) may be obtained by liberating the compound of formula (I) from one of their functional derivatives by treatment with solvolytic or hydrogenolytic agents.

Preferred feedstocks for solvolysis or hydrogenolysis are feedstocks which: those which are in accordance with formula (I), but which contain a corresponding protected amino and/or hydroxyl group (instead of one or more free amino and/or hydroxyl groups), preferably those which carry an amino protecting group (instead of the H atom bound to the N atom), in particular those which carry an R '-N group (where R' represents an amino protecting group) (instead of the HN group) and/or those which carry a hydroxyl protecting group (instead of the H atom of a hydroxyl group), for example those which are in accordance with formula (I), but which carry a-COOR "group (where R" represents a hydroxyl protecting group) (instead of the-COOH group).

It is also possible for a plurality of identical or different protected amino and/or hydroxyl groups to be present in the starting molecule. If protecting groups different from each other are present, they can be selectively cleaved off in many cases.

The term "amino protecting group" is a known general term and refers to a group that: suitable for protecting (blocking) the amino group from chemical reactions, but which can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Representative of such groups are in particular: unsubstituted or substituted acyl, aryl, aralkoxymethyl or aralkyl. Their type and size are not critical since the amino protecting group is removed after the desired reaction (or reaction sequence); however, those having 1 to 20, in particular 1 to 8, carbon atoms are preferred. The term "acyl" is to be understood in its broadest sense in relation to the process of the present invention. It includes acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic or sulfonic acids and in particular alkoxy-carbonyl, aryloxycarbonyl, especially aralkoxycarbonyl groups. Examples of such acyl groups are: alkanoyl groups such as acetyl, propionyl, and butyryl; aralkanoyl such as phenylacetyl; aroyl groups such as benzoyl and tolyl (tolyl); aryloxyalkanoyl such as POA; alkoxycarbonyl such as methoxy-carbonyl, ethoxycarbonyl, 2,2, 2-trichloroethoxycarbonyl, BOC (tert-butoxy-carbonyl) and 2-iodoethoxycarbonyl; aralkoxycarbonyl such as CBZ ("carbobenzoxy"), 4-methoxybenzyloxycarbonyl and FMOC; and aryl-sulfonyl groups such as Mtr. Preferred amino protecting groups are BOC and Mtr, as well as CBZ, Fmoc, benzyl and acetyl.

The term "hydroxy protecting group" is also a known generic term and refers to groups that: suitable for protecting hydroxyl groups from chemical reactions, but can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Representative of such groups are the unsubstituted or substituted aryl, aralkyl or acyl groups mentioned above, as well as alkyl groups. The nature and size of the hydroxyl protecting groups is not critical since they are removed after the desired chemical reaction or reaction sequence; preference is given to radicals having from 1 to 20, in particular from 1 to 10, carbon atoms. Examples of hydroxyl-protecting groups are, inter alia: benzyl, 4-methoxybenzyl, p-nitro-benzoyl, p-toluenesulfonyl, tert-butyl and acetyl, of which benzyl and tert-butyl are particularly preferred.

The term "solvate of a compound" is used to denote an adduct (adduct) of an inert solvent molecule on a compound, which is formed due to their mutual attraction. Solvates are, for example, mono-or dihydrate or alcoholates (alcoholates).

Depending on the protecting group used, for example a strong acid, advantageously TFA or perchloric acid, may be used, but it is also possible to liberate the compounds of formula (I) from their functional derivatives using other strong inorganic acids, such as hydrochloric acid or sulfuric acid, strong organic carboxylic acids, such as trichloroacetic acid, or sulfonic acids, such as benzenesulfonic acid or p-toluenesulfonic acid. Other inert solvents may be present, but are not always necessary. Suitable inert solvents are preferably organic solvents, for example: carboxylic acids (such as acetic acid), ethers (such as THF or dioxane), amides (such as DMF), halogenated hydrocarbons (such as DCM), and alcohols (such as methanol, ethanol or isopropanol) and water. Mixtures of the above solvents are also suitable. An excess of TFA is preferably used without addition of further solvent, and the perchloric acid is preferably used in the form of a mixture of acetic acid and 70% perchloric acid in a ratio of 9: 1. The reaction temperature for cleavage is advantageously between about 0 and about 50 ℃, preferably between 15-30 ℃ (room temperature).

For example, the BOC, OBut, and Mtr groups can be cleaved off preferably at 15-30 ℃ using TFA in DCM or using about 3-5N HCl in dioxane, and the FMOC groups can be cleaved off at 15-30 ℃ using about 5-50% solutions of dimethylamine, diethylamine, or piperidine in DMF.

Protecting groups which can be removed hydrogenolytically (e.g. the liberation of CBZ, benzyl or amidino from its oxadiazole derivative) can be cleaved off, for example, by treatment with hydrogen in the presence of a catalyst (e.g. a noble metal catalyst such as palladium, advantageously on a support such as carbon). Suitable solvents here are those mentioned above, in particular, for example, alcohols (such as methanol or ethanol) or amides (such as DMF). The hydrogenolysis is generally carried out at a temperature of between about 0 and 100 ℃ and a pressure of between about 1 and 200 bar, preferably between 20 and 30 ℃ and 1 to 10 bar. For example, hydrogenolysis of the CBZ group is carried out well in methanol at 5-10% Pd/C, or using ammonium formate (instead of hydrogen) in methanol/DMF at Pd/C, at 20-30 ℃.

Examples of suitable inert solvents are: hydrocarbons (such as hexane, petroleum ether, benzene, toluene, or xylene); chlorinated hydrocarbons (such as trichloroethylene, 1, 2-dichloroethane, tetrachloromethane, trifluoromethylbenzene, chloroform or DCM); alcohols (such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol); ethers (such as diethyl ether, diisopropyl ether, Tetrahydrofuran (THF), or dioxane); glycol ethers (such as ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, or ethylene glycol dimethyl ether (diglyme)); ketones (such as acetone or butanone); amides such as acetamide, dimethylacetamide, N-methylpyrrolidone (NMP) or dimethyl-formamide (DMF); nitriles (such as acetonitrile); sulfoxides (such as dimethyl sulfoxide (DMSO)); carbon disulfide; carboxylic acids (such as formic acid or acetic acid); nitro compounds (such as nitromethane or nitrobenzene); esters (such as EtOAc); or mixtures of said solvents.

The esters can be saponified, for example, using LiOH, NaOH or KOH in water, water/THF/ethanol or water/dioxane, at temperatures between 0-100 ℃. In addition, the ester can be hydrolyzed, for example, using acetic acid, TFA, or HCL.

Furthermore, the free amino group can be acylated using an acid chloride or anhydride, or alkylated using an unsubstituted or substituted alkyl halide, or reacted with CH in a conventional manner₃-C (= NH) -OEt reaction, advantageously in an inert solvent such as DCM or THF, and/or in the presence of a base such as triethylamine or pyridine, at a temperature of-60 ℃ to +30 ℃.

In the present specification, the term leaving group preferably denotes Cl, Br, I or a reactively modifiable OH group, e.g. an activated ester, an imidazole anion (imidazolide) or an alkylsulfonyloxy group having 1 to 6 carbon atoms (preferably a methylsulfonyloxy or trifluoromethylsulfonyloxy group) or an arylsulfonyloxy group having 6 to 10 carbon atoms (preferably a phenylsulfonyloxy or p-toluenesulfonyloxy group).

Such groups are described in the literature for activating carboxyl groups in typical acylation reactions (for example in standard works such as Houben-Weyl, Methoden der organischen Chemistry [ Methods of Organic Chemistry ], Georg-Thieme-Verlag, Stuttgart).

The activated ester is advantageously formed in situ, for example by addition of HOBt or N-hydroxysuccinimide.

The term "pharmaceutically acceptable derivatives" is used to indicate, for example, salts of the compounds of formula I and so-called prodrug compounds.

The term "prodrug derivative" is used to denote a compound of formula I which has been modified, for example, by an alkyl or acyl group, a sugar such as glucuronide, or an oligopeptide, and which will be rapidly cleaved in the organism to form the active compound.

These also include biodegradable polymer derivatives of the compounds according to the invention, which are described, for example, in int. j. Pharm.11561-67 (1995).

Metabolites of compounds of formula (I) and related formulae are also part of the invention. The metabolite of the compound of formula (I) may be, for example, wherein Y¹And/or Y²A compound of formula (I) which is a glucuronide. Examples of such metabolites are for example compounds of formula (Ik).

The metabolite of the compounds of formula (I) and related formulae may also be the NY bound therein¹-W₃Compounds of formula (I) and related formulae which have been cleaved, e.g. formula (I)^M) As shown.

WhereinG¹、G²、R^a、R^b、R¹、W¹、X¹、X²、W²、Y¹As defined above. Examples of such metabolites are the compounds M1 and M2 mentioned in table I below.

The metabolites of the compounds of formula (I) and related formulae may also be compounds of formula (I) and related formulae in which one or more of the nitrogen atoms have been oxidized. The compounds of examples 101, 102 and 103 represent examples of such metabolites.

The metabolites of the compounds of formula (I) and related formulae may also be compounds of formula (I) and related formulae in which one or more aromatic systems, such as phenyl rings, are oxidized. Such metabolites may be represented by the following formula (I)^M2) Definition of

Wherein G is³、G⁴、G⁵May independently of one another represent OH, t is 0, 1, 2 or 3, preferably 1 or 2, G¹、G²、R^a、R^b、R¹、X¹、X²、Y¹、Y²、Z¹、Z²、W¹、W²、W³And W⁴As defined above. Examples of such metabolites are the following compounds M3, M6 and M9:

。

the present invention includes compounds of formula (I) and related formulae, alone or in combination with one or more metabolites thereof.

Formula (I) also includes mixtures of compounds of formula I, for example mixtures of 2 diastereomers (e.g., in a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:100, or 1: 1000).

These are particularly preferably mixtures of stereoisomeric compounds.

In another preferred embodiment, the compounds of formula (I) exhibit an affinity for S₁P₁Superiority of the receptor over S₁P₃Selectivity of the receptor, based on an IC50 value of more than 20-fold, preferably more than 50-fold, more preferably more than 100-fold, even more preferably more than 1000-fold, as determined in the examples described below.

Preferred compounds of the invention have high oral bioavailability and/or low clearance.

In another preferred embodiment, the compounds of the present invention exhibit activity against S₁P₁Superiority of the receptor over S₁P₃Receptor selectivity, based on an IC50 value of more than 20-fold, preferably more than 50-fold, more preferably more than 100-fold, even more preferably more than 1000-fold as determined in the examples below and the invention has high oral bioavailability and/or low clearance.

Preferably, the compounds of the present invention are selected from the following examples 1-96 and pharmaceutically acceptable derivatives, solvates, salts and stereoisomers thereof, including mixtures thereof in all ratios:

example numbering	Formula (II)
		1
2
		3
4
		5
6
		7
8
		9
10
		11
12
		13
14
		15
16
		17
18
		19
20
		21
22
		23
24
		25
26
		27
28
		29
30
		31
32
		33
34
		35
36
		37
38
		39
40
		41
42
		43
44
		45
46
		47
48
		49
50
		51
52
		53
54
		55
56
		57
58
		59
60
		61
62
		63
64
		65
66
		67
68
		69
70
		71
72
		73
74
		75
76
		77
78
		79
80
		81
82
		83
84
		85
86
		87
88
		89
90
		91
92
		93
94
		95
96
		97
98
		99
100
		101
102
		103
M1
		M2

Formula (I) also includes optically active forms (stereoisomers), enantiomers, racemates, diastereomers and hydrates, salts and solvates of these compounds.

In a particular embodiment, when 2 or more chiral centers are present, the compound of formula (I) is obtained as one diastereomer.

"diastereomer" means that each chiral center present in the compound of formula (I) is defined relative to the other chiral centers.

For all groups and markers (indices) that occur more than 1 time within the same chemical structure, their meanings are independent of each other.

In the above and in the following, the radical or the parameter R^a、R^b、X¹、X²、Y¹、Y²、G¹、A、A、Z¹、Z²、W¹、W²、W³、W⁴Have the meaning indicated under formula (I) and subformulae unless explicitly stated otherwise.

Cycloalkyl is a cyclic alkyl group containing from 3 to 12 carbon atoms. Cycloalkyl preferably denotes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.

Cycloalkylalkylene is cycloalkyl bonded to the rest of the molecule through a carbon chain and having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms. Cycloalkylalkylene preferably denotes cyclopropylmethylene, cyclobutylmethylene, cyclopentylmethylene, cyclohexylmethylene or cycloheptylmethylene.

R^aIs preferably-CH₃、-CH₂CH₃、-CF₃、-CHF₂，

R^bPreferably is-CF₃or-CH₂OCH₃。

In one embodiment, the invention provides wherein R^aAnd R^bOne is-CF₃or-CH₂OCH₃preferably-CF₃A compound of formula (I).

Halogen is preferably F, Cl or Br, in particular F or Cl.

R¹Preferably F or OA (wherein A is as defined above), in particular F or-OCH₃，

A group in the formula (I)Examples of (a) are selected from the following groups:

the compounds of formula (I) may have one or more chiral centers and may therefore exist in a variety of stereoisomeric forms. Formula (I) covers all these forms.

In addition, by Methods such as those described in the literature (for example in standard works, such as Houben-Weyl, Methoden der organischen Chemistry [ Methods of Organic Chemistry ]]The process known per se, described in Georg-Thieme-Verlag, Stuttgart), under reaction conditions known and suitable for the reaction, the compounds of the formula (I) and the starting materials for their preparation. For all protection and deprotection methods see Philip J. Kocienski,') "Protecting Groups", Georg Thieme Verlag Stuttgart, New York, 1994 and Theodora W.Greene and Peter G.M.Wuts"Protective Groups in Organic Synthesis", Wiley Interscience, 3 rd edition 1999.

Variants known per se but not mentioned here in more detail can also be used here.

The starting materials may also be formed in situ, if desired, so that they are not isolated from the reaction mixture but immediately converted further into the compounds of the formula (I).

The starting compounds for preparing the compounds of the formula (I) are generally known. However, if they are not novel, they can be prepared by methods known per se.

The reaction is preferably carried out in an inert solvent.

Examples of suitable inert solvents are: hydrocarbons (such as hexane, petroleum ether, benzene, toluene, or xylene); chlorinated hydrocarbons (such as trichloroethylene, 1, 2-dichloroethane, tetrachloromethane, chloroform or DCM); alcohols (such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol); ethers (such as diethyl ether, diisopropyl ether, thf (thf), or dioxane); glycol ethers (such as ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, or ethylene glycol dimethyl ether (diglyme)); ketones (such as acetone or butanone); amides (such as acetamide, dimethylacetamide or Dimethylformamide (DMF)); nitriles (such as acetonitrile); sulfoxides (such as dimethyl sulfoxide (DMSO)); carbon disulfide; carboxylic acids (such as formic acid or acetic acid); nitro compounds (such as nitromethane or nitrobenzene); esters (such as EtOAc); or mixtures of said solvents.

Pharmaceutical salts and other forms

The compounds of formula (I) may be used in their final non-salt form. In another aspect, the invention also relates to the use of these compounds in the form of their pharmaceutically acceptable salts, which can be derived from various organic and inorganic acids and bases by procedures known in the art. Pharmaceutically acceptable salt forms of the compounds of formula I are generally prepared by conventional methods. If a compound of formula I contains an acidic centre (such as a carboxyl group), one of its suitable salts may be formed by reacting the compound with a suitable base to form the corresponding base addition salt. Such bases are, for example, alkali metal hydroxides, including potassium hydroxide and sodium hydroxide; alkaline earth metal hydroxides such as magnesium hydroxide and calcium hydroxide; and various organic bases such as piperidine, diethanolamine and N-methyl-glucosamine (meglumine), benzathine (benzathine), choline, diethanolamine, ethylenediamine, benzphetamine, diethylamine, piperazine, lysine, L-arginine, ammonia, triethanolamine, betaine, ethanolamine, morpholine and tromethamine. In the case of certain compounds of formula I containing a basic center, acid addition salts may be formed by treating these compounds with: pharmaceutically acceptable organic and inorganic acids, for example hydrogen halides (such as hydrogen chloride or hydrogen bromide), other inorganic acids and their corresponding salts (such as sulfates, nitrates or phosphates, etc.), and alkylsulfonates and monoarylsulfonates (such as methanesulfonates, ethanesulfonates, toluenesulfonates and benzenesulfonates), and other organic acids and their corresponding salts (such as carbonates, acetates, trifluoroacetates, tartrates, maleates, succinates, citrates, benzoates, salicylates, ascorbates, etc.). Thus, pharmaceutically acceptable acid addition salts of compounds of formula I include the following: acetate, adipate, alginate, aspartate, benzoate, benzene-sulfonate (benzenesulfonate), bisulfate, bisulfite (bisulphite), bromide, camphorate, camphorsulfonate, caprate, caprylate, chloride, chlorobenzoate, citrate, cyclamate, cinnamate, digluconate, dihydrogen phosphate, dinitrobenzoate, dodecyl-sulfate, ethanesulfonate, formate, glycolate, fumarate, galactarate (galactarate) (from mucic acid), galacturonate, glucoheptonate (glucoheptonate), gluconate, glutamate, glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isethionate, dihydrocarb (bisulfite), bromide, camphor-oxide, camphor-sulfate, caprylate, fumarate, caprylate, 2-hydroxyethanesulfonate, iodide, sodium hydrogen succinate, sodium hydrogen, Isobutyrate, lactate, lactobionate, malate, maleate, malonate, mandelate, metaphosphate, methanesulfonate, methylbenzoate, monohydrogenphosphate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, oleate, palmitate, pectate, persulfate, phenylacetate, 3-phenylpropionate, phosphate, phosphonate, phthalate, but this does not represent a limitation. Both types of salts can be formed or interconverted preferably using ion exchange resin techniques.

Further, base salts of compounds of formula I include aluminum, ammonium, calcium, copper, iron (III), iron (II), lithium, magnesium, manganese (III), manganese (II), potassium, sodium, and zinc salts, although this is not intended to represent a limitation. Among the above salts, preferred are: an ammonium salt; alkali metal sodium and potassium salts; and alkaline earth metal calcium and magnesium salts. Salts of compounds of formula I derived from pharmaceutically acceptable organic non-toxic bases include salts of: primary, secondary and tertiary amines, substituted amines (also including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, chloroprocaine, choline, N' -dibenzylethylenediamine (benzathine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, histidine, hydrabamine, isopropylamine, lidocaine, lysine, meglumine (N-methyl-D-glucamine), morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethanolamine, triethylamine, trimethylamine, tripropylamine, and tris (hydroxymethyl) methylamine (tromethamine), this is not intended to be limiting.

Containing alkaline N₂The compounds of formula I of the present invention of the group may be quaternized with the following agents: such as (C1-C4) alkyl halides, e.g., methyl, ethyl, isopropyl and tert-butyl chlorides, bromides and iodides; di (C1-C4) alkyl sulfates such as dimethyl, diethyl, and diamyl sulfates; (C10-C18) alkyl halides such as decyl, dodecyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; and aryl (C1-C4) alkyl halides, such as benzyl chloride and phenethyl bromide. Using such salts, water-soluble and oil-soluble compounds of formula I can be prepared.

Preferred pharmaceutical salts mentioned above include: acetate, trifluoroacetate, benzenesulfonate, citrate, fumarate, gluconate, hemisuccinate, hippurate, hydrochloride, hydrobromide, isethionate, mandelate, meglumine, nitrate, oleate, phosphonate, pivalate, sodium phosphate, stearate, sulfate, sulfosalicylate, tartrate, thiomalate, tosylate and tromethamine, although this is not intended to represent a limitation.

An acid addition salt of a basic compound of formula (I) is prepared as follows: in a conventional manner, the free base form is contacted with a sufficient amount of the desired acid to form a salt. The free base can be regenerated as follows: the salt form is contacted with a base in a conventional manner and the free base is isolated. The free base form differs somewhat from its corresponding salt form in some respects with respect to certain physical properties, such as solubility in polar solvents, but otherwise the salts are comparable to their respective free base forms for the purposes of the present invention.

As indicated, pharmaceutically acceptable base addition salts of compounds of formula I are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Preferred metals are sodium, potassium, magnesium and calcium. Preferred organic amines are N, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D-glucosamine and procaine.

Base addition salts of acidic compounds of formula I are prepared as follows: in a conventional manner, the free acid form is contacted with a sufficient amount of the desired base to form a salt. The free acid can be regenerated as follows: the salt form is contacted with an acid in a conventional manner and the free acid is isolated. The free acid form differs somewhat from its corresponding salt form in some respects with respect to certain physical properties, such as solubility in polar solvents, but otherwise the salts are comparable to their respective free acid forms for the purposes of the present invention.

If the compound of formula (I) contains more than one group capable of forming such pharmaceutically acceptable salts, then formula I also includes multiple salts. Typical multi-salt forms include, for example, bitartrate, diacetate, hydrogen fumarate, meglumine, diphosphate, disodium, and trihydrochloride, but this is not intended to represent a limitation.

As can be seen from the above, the term "pharmaceutically acceptable salt" in this respect is used to indicate an active ingredient which is: comprising a compound of formula I in one of its salt forms, which salt forms impart improved pharmacokinetic properties to the active ingredient, in particular compared to the free form of the active ingredient or any other salt form of the active ingredient used before. The pharmaceutically acceptable salt form of the active ingredient may also provide the active ingredient for the first time with desirable pharmacokinetic properties not previously present and may even have a positive effect on the pharmacodynamics of the active ingredient in its in vivo therapeutic effect.

Due to their molecular structure, the compounds of formula (I) may be chiral and may therefore exist in different enantiomeric forms. Thus, they may be present in racemic form or in optically active form.

Since the pharmaceutical activity of racemates or stereoisomers of the compounds according to the invention may differ, it may be desirable to use enantiomers. In these cases, the end products or even intermediates can be separated into the enantiomeric compounds by chemical or physical measures known to the person skilled in the art, or even used as such in the synthesis.

In the case of racemic amines, diastereomers are formed from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as in the (R) and (S) forms: tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, a suitable N-protected amino acid (e.g. N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulphonic acids. Also advantageous is chromatographic enantiomeric resolution with the aid of optically active resolving agents (e.g. dinitrobenzoylphenylglycine, cellulose triacetate or other derivatives of carbohydrates or chirally derivatized methacrylate polymers fixed on silica gel). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile (for example in a ratio of 82:15: 3).

The invention furthermore relates to the use of compounds of the formula I and related formulae in combination with the following agents: at least one further pharmaceutically active ingredient, preferably a drug for the treatment of multiple sclerosis such as cladribine, or other co-agents such as interferons, e.g. pegylated or non-pegylated interferons, preferably interferon beta, and/or compounds that improve vascular function, or immunomodulators such as fingolimod; cyclosporin, rapamycin or ascomycin or their immunosuppressive analogs such as cyclosporin A, cyclosporin G, FK-506, ABT-281, ASM981, rapamycin, 40-O- (2-hydroxy) ethyl-rapamycin, and the like; a corticosteroid; cyclophosphamide; azathioprine; methotrexate; leflunomide; mizoribine; mycophenolic acid (mycophenolic add); mycophenolate mofetil (mycophenolate mofetil); 15-deoxyspergualin; diflucortolone valerate; difluprednate; alclometasone dipropionate; amcinonide; amsacrine; asparaginase enzyme; azathioprine; basiliximab; beclomethasone dipropionate; betamethasone; betamethasone acetate; betamethasone dipropionate; betamethasone sodium phosphate (betamethasone phosphate solution); betamethasone valerate; budesonide; captopril; mechlorethamine hydrochloride; cladribine; clobetasol propionate; cortisone acetate; (ii) a kovar; cyclophosphamide; cytarabine; (ii) daclizumab; dactinomycin; (ii) donepezil; desoximetasone; dexamethasone; dexamethasone acetate; dexamethasone isonicotinic acid; dexamethasone sodium metasulfobenzoate (dexamethasone metasulfonbenzoate sodium); dexamethasone phosphate; dexamethasone tert-butylacetate; acetic acid dichloropine; doxorubicin hydrochloride; epirubicin hydrochloride; (ii) chlorofluoronenide; fludrocortisone acetate; fluoro-hydrogen shrinkage; flumethasone pivalate; flunisolide; fluocinolone acetonide; fluocinonide; fluocortolone; fluocortolone caproate; flucolone pivalate; fluorometholone; fluprednidene acetate; fluticasone propionate; gemcitabine hydrochloride; halcinonide; hydrocortisone, hydrocortisone acetate, hydrocortisone butyrate, hydrocortisone hemisuccinate; melphalan; methylprednisolone; mercaptopurine; methylprednisolone; methylprednisolone acetate; methylprednisolone hemisuccinate; misoprostol; muromab-cd 3; mycophenolate mofetil; (ii) paramethasone acetate; pinazoline, prednisolone; prednisolone acetate; prednisolone caproate; prednisolone sodium metasulfonoate sodium (prednisolone metasulfonoate sodium); prednisolone sodium phosphate (prednisolone phosphate sodium); prednisone; prednisolone; rifampin; rifampicin sodium (rifampicine sodique); tacrolimus; teriflunomide; thalidomide; thiotepa; tixocortole pivalate; triamcinolone acetonide; triamcinolone hemisuccinate; triamcinolone acetonide; triamcinolone diacetate; triamcinolone acetonide hexa; immunosuppressive monoclonal antibodies, e.g., monoclonal antibodies directed against leukocyte receptors, e.g., MHC, CD2, CD3, CD4, CD7, CD25, CD28, B7, CD40, CD45, or CD58 or ligands thereof; or other immunomodulatory compounds, such as CTLA41g or other adhesion molecule inhibitors, such as mabs or low molecular weight inhibitors, including selectin antagonists and VLA-4 antagonists. One preferred composition contains cyclosporin A, FK506, rapamycin or 40- (2-hydroxy) ethyl-rapamycin and fingolimod. These other drugs (such as interferon beta) can be administered concomitantly or sequentially, for example by subcutaneous, intramuscular or oral routes.

These compositions are useful as medicaments in human and veterinary medicine.

The pharmaceutical preparations may be administered in the form of dosage units, each containing a predetermined amount of the active ingredient. Such units may contain, for example, from 0.5 mg to 1 g, preferably from 1 mg to 700 mg, particularly preferably from 5 mg to 100 mg, of a compound according to the invention, depending on: the disease condition to be treated, the method of administration and the age, weight and condition of the patient, or the pharmaceutical preparation, may be administered in the form of dosage units, each containing a predetermined amount of the active ingredient. Preferred dosage unit formulations are those wherein: which comprises the above-mentioned daily dose or divided dose of the active ingredient, or their respective fractions. In addition, such pharmaceutical preparations may be prepared using methods generally known in the pharmaceutical art.

The pharmaceutical formulation may be adapted for administration via any desired suitable method, for example by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or intradermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) methods. Such formulations may be prepared using all methods known in the pharmaceutical art, for example by combining the active ingredient with an excipient(s) or adjuvant(s).

Pharmaceutical formulations adapted for oral administration may be administered as separate units, for example, capsules or tablets; powder or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or foam foods; or an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.

Thus, for example, in the case of oral administration in the form of tablets or capsules, the active ingredient component may be combined with an oral, non-toxic and pharmaceutically acceptable inert excipient (e.g., ethanol, glycerol, water, and the like). Powders were prepared as follows: the compound is ground to a suitable fineness and mixed with a pharmaceutical excipient (e.g., an edible carbohydrate such as starch or mannitol) that is ground in a similar manner. Flavors, preservatives, dispersants, and dyes may be similarly present.

Capsules were prepared by preparing a powder mixture as described above and filling it into shaped gelatin shells. Glidants and lubricants in solid form (e.g., highly dispersed silicic acid, talc, magnesium stearate, calcium stearate or polyethylene glycol) may be added to the powder mixture prior to the filling operation. Disintegrating or solubilizing agents, such as agar-agar, calcium carbonate or sodium carbonate, may also be added to enhance the availability of the drug after administration of the capsule.

In addition, if desired or necessary, suitable binders, lubricants and disintegrants and dyes may also be incorporated into the mixture. Suitable binders include: starches, gelatin, natural sugars (e.g., glucose or beta-lactose), sweeteners made from corn, natural and synthetic gums (e.g., acacia, tragacanth or sodium alginate), carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include: sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. Tablets were formulated as follows: for example, a powder mixture is prepared, the mixture is granulated or dry-pressed (dry-pressing), a lubricant and a disintegrant are added, and the whole mixture is compressed to obtain a tablet. A powder mixture was prepared as follows: the compound, comminuted in a suitable manner, is mixed with a diluent or base as described above, and optionally with: binders (e.g., carboxymethylcellulose, alginates, gelatin, or polyvinylpyrrolidone), dissolution retardants (e.g., paraffin), absorption enhancers (e.g., quaternary salts), and/or adsorbents (e.g., bentonite, kaolin, or dicalcium phosphate). The powder mixture can be granulated as follows: it is wetted with a binder (e.g., syrup, starch paste, acadia (acadia) mucilage, or a solution of cellulose or polymeric material) and forced through a screen. As an alternative to granulation, the powder mixture may be run through a tablet press to produce a non-uniformly shaped mass that is comminuted to form granules. The granules can be lubricated to prevent sticking to the tablet mold by the addition of stearic acid, stearate, talc or mineral oil. The lubricated mixture is then compressed to obtain tablets. The active ingredient may also be combined with free-flowing inert excipients and then directly compressed to give tablets without the need for granulation or dry compression steps. There may be a protective layer consisting of a shellac sealing layer, which may be transparent or opaque, a layer of sugar or polymer material and a gloss layer of wax. Dyes may be added to these coatings to enable differentiation of different dosage units.

Oral liquids (e.g., solutions, syrups and elixirs) may be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared by dissolving the compound in an aqueous solution containing a suitable flavoring agent, while elixirs are prepared using a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound in a non-toxic vehicle. It is also possible to add: solubilizers and emulsifiers (e.g., ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers), preservatives, flavoring additives (e.g., peppermint oil or natural sweeteners or saccharin), or other artificial sweeteners, and the like.

Dosage unit formulations for oral administration may be encapsulated in microcapsules, if desired. The formulations may also be prepared in a prolonged or delayed release manner, for example, by coating or embedding the microparticles with polymers, waxes, and the like.

The compounds of formula (I) and salts, solvates and physiologically functional derivatives thereof as well as other active ingredients may also be administered in the form of liposome delivery systems, e.g., small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids (e.g., cholesterol, stearamide, or phosphatidylcholine).

The compounds of formula (I) and salts, solvates and physiologically functional derivatives thereof as well as other active ingredients may also be delivered using monoclonal antibodies as single carriers to which the compound molecules are coupled. The compounds may also be conjugated to soluble polymers used as targeted drug carriers. Such polymers may include: polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol (polyhydroxypropylamidophenol), polyhydroxyethylaspartamidophenol (polyhydroxyethylaminophenol), or polyethyleneoxide polylysine (which is substituted with palmitoyl). The compounds may also be coupled to a class of biodegradable polymers suitable for achieving controlled release of a drug, such as polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, and crosslinked or amphiphilic hydrogel block copolymers.

Pharmaceutical formulations adapted for transdermal administration may be administered as a stand-alone plaster for prolonged intimate contact with the epidermis of the recipient. Thus, for example, active ingredients may be delivered from plasters by iontophoresis, which is described as a general term in Pharmaceutical Research, 3(6), 318 (1986).

Pharmaceutical compounds suitable for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.

For the treatment of the eye or other external tissues (e.g. mouth and skin), the formulation is preferably applied as a topical ointment or cream. In the case of formulation to produce an ointment, the active ingredient may be used with a paraffinic (parafinic) or with a water-miscible cream base. Alternatively, the active ingredient may be formulated with an oil-in-water cream base or a water-in-oil base to give a cream.

Pharmaceutical formulations suitable for topical administration to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent.

Pharmaceutical formulations suitable for topical administration in the mouth include lozenges, pastilles and mouthwashes.

Pharmaceutical preparations suitable for rectal administration may be administered in the form of suppositories or enemas.

Pharmaceutical formulations suitable for nasal administration, wherein the carrier material is a solid, include coarse powders having a particle size, for example, in the range of 20-500 microns, which are administered by inhalation from the nose, that is, by rapid inhalation through the nasal passage from a container containing the powder held in close proximity to the nose. Suitable formulations for administration as a nasal spray or nasal drops (with a liquid as the carrier material) include solutions of the active ingredient in water or oil.

Pharmaceutical formulations suitable for administration by inhalation include finely divided particulate powders or mists which may be generated by various types of pressurised aerosol-containing dispensers, nebulisers or inhalers.

Pharmaceutical formulations suitable for vaginal administration may be administered as pessaries, tampons, creams, gels, pastes, foams or spray formulations.

Pharmaceutical formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions containing antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the recipient to be treated; and aqueous and non-aqueous sterile suspensions which may contain a suspending vehicle and a thickening agent. The formulations may be administered in single-or multi-dose containers, for example sealed ampoules and vials, and stored in a freeze-dried (lyophilised) state, so that only the addition of a sterile carrier liquid, for example water for injections, is required immediately prior to use.

Injectable solutions and suspensions, prepared according to the prescription, can be prepared from sterile powders, granules and tablets.

It goes without saying that, in addition to the components specifically mentioned above, the formulations may also comprise other agents common in the art with respect to the specific formulation type; thus, for example, formulations suitable for oral administration may contain flavoring agents.

The therapeutically effective amount of the compound of formula I and other active ingredients will depend on a number of factors including, for example, the age and weight of the animal, the precise condition being treated and its severity, the nature of the formulation and the method of administration, and will ultimately be determined by the attending physician or veterinarian. However, an effective amount of the compound will generally be in the range of from 0.1 to 100 mg/kg body weight of the recipient (mammal) per day, with an amount in the range of from 1 to 10 mg/kg body weight per day being particularly typical. Thus, for an adult mammal weighing 70 kg, the actual amount per day is typically between 70-700 mg, where the amount may be administered as a single dose per day, or often in a series of divided doses per day (e.g., 2, 3, 4, 5, or 6), such that the total daily dose is the same. An effective amount of a salt or solvate or physiologically functional derivative thereof may be determined as a fraction of the effective amount of the compound itself.

The invention further relates to a method for treating a subject suffering from a sphingosine 1-phosphate related disorder, said method comprising: administering to the subject an effective amount of a compound of formula (I). The present invention preferably relates to such a method, wherein said sphingosine 1-phosphate-1 related disorder is an autoimmune disorder or disease associated with an overactive immune response.

The invention further relates to a method of treating a subject suffering from an immunoregulatory abnormality, the method comprising: administering to the subject a compound of formula (I) in an amount effective to treat the immunoregulatory abnormality. The present invention preferably relates to such a method, wherein said immunoregulatory abnormality is an autoimmune disease or a chronic inflammatory disease selected from the group consisting of: amyotrophic Lateral Sclerosis (ALS), systemic lupus erythematosus, chronic rheumatoid arthritis, type I diabetes, inflammatory bowel disease, biliary cirrhosis, uveitis, multiple sclerosis, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis, autoimmune myositis, Wegener's granulomatosis, ichthyosis, Graves ' ophthalmopathy, and asthma. The invention further relates to such a method, wherein the immunoregulatory abnormality is bone marrow or organ transplant rejection or graft versus host disease. The invention further relates to such a method, wherein the immunoregulatory abnormality is selected from the group consisting of: transplantation of organ or tissue, graft-versus-host disease resulting from transplantation, autoimmune syndromes including rheumatoid arthritis, systemic lupus erythematosus, hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, uveitis, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, autoimmune diseases following infection (including rheumatic fever and glomerulonephritis following infection), inflammatory and hyperproliferative skin diseases, psoriasis, atopic dermatitis, contact dermatitis, eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitis, erythema, cutaneous eosinophilia, lupus erythematosus, acne, alopecia areata, keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, Keratitis, herpetic keratitis, keratoconus, corneal epithelial dystrophy, leukoplakia, ocular pemphigus, moren's ulcer, scleritis, Graves ' opthalmopathy, voguet-salix minor-quardt syndrome, sarcoidosis, pollen allergy, reversible obstructive airways disease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma, dust asthma, chronic or refractory asthma, late asthma and airway hyperreactivity, bronchitis, gastric ulcer, vascular damage caused by ischemic disease and thrombosis, ischemic bowel disease, inflammatory bowel disease, necrotizing enterocolitis, small intestine injury associated with thermal burn, abdominal disease, proctitis, eosinophilic gastroenteritis, mastocytosis, crohn's disease, ulcerative colitis, migraine headache, Rhinitis, eczema, interstitial nephritis, goodpasture's syndrome, hemolytic uremic syndrome, diabetic nephropathy, polymyositis, guillain-barre syndrome, meniere's disease, polyneuritis, mononeuritis, radiculopathy, hyperthyroidism, Barceltis disease, simple red cell aplasia, aplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia, erythropoiesis inability, osteoporosis, sarcoidosis, fibrotic lung, idiopathic interstitial pneumonia, dermatomyositis, leukoplakia vulgaris, ichthyosis vulgaris, photoallergic sensitivity (photoallergic sensitivity), cutaneous T-cell lymphoma, chronic lymphocytic leukemia, arteriosclerosis, atherosclerosis, diabetes, chronic lymphocytic leukemia, chronic myelogenous, Aortosis syndrome, polyarteritis nodosa, non-inflammatory cardiomyopathy, scleroderma, wegener's necrotic granuloma, sjogren's syndrome, obesity, eosinophilic fasciitis (eosinophilic fasciitis), gingival injury, periodontal tissue injury, alveolar bone injury, cementum injury, glomerulonephritis, male or senile alopecia (by preventing or providing hair loss and/or promoting hair germination and hair growth), muscular dystrophy, pyoderma and sezary's syndrome, addison's disease, organ ischemia-reperfusion injury following preservation, transplantation or ischemic disease, endotoxic shock, pseudomembranous colitis, colitis caused by drugs or radiation, ischemic acute renal insufficiency, chronic renal insufficiency, toxemia caused by lung-oxygen or drugs, lung cancer, renal cancer, Emphysema, cataract, lung iron deposition, retinitis pigmentosa, age-related macular degeneration, vitreous scarring, corneal alkali burn, dermatitis erythema multiforme (dermatitis erythrematosis multiforme), linear IgA ballous dematitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution, aging, carcinogenesis (carcinogenesis), cancer metastasis and baropathy, diseases caused by histamine or leukotriene 4 release, liver function Behcet's disease, autoimmune hepatitis, primary biliary cirrhosis, sclerosing cholangitis, partial hepatectomy, acute hepatic necrosis, necrosis caused by toxins, viral hepatitis, shock or hypoxia, B-viral hepatitis, non-A/non-B hepatitis, hepatic cirrhosis, alcoholic hepatic cirrhosis, liver failure, hair loss, delayed failure, chronic liver failure, and/kidney disease, "Chronic" liver failure, potentiation of chemotherapy, cytomegalovirus infection, HCMV infection, AIDS, cancer, Alzheimer's disease, trauma, and chronic bacterial infection.

In a preferred embodiment, the compounds of the invention are useful for the treatment of multiple sclerosis, including relapsing-remitting multiple sclerosis.

Preferred compounds of formula (I) exhibit the followingBinding measured according to an embodiment of the invention S1P₁Ki of the receptor: less than about 10 μ M, preferably less than about 5 μ M, more preferably less than about 1 μ M, and even more preferably less than about 0,1 μ M. Most preferably, the compound of formula (I) exhibits less than 0.01. mu.M binding to S1P₁Ki of (1).

In the following, the invention will be illustrated by means of some examples, which should not be construed as limiting the scope of the invention.

Examples

The oxadiazole compounds according to formula (I) can be prepared from readily available starting materials by several synthetic methods using solution phase and solid phase chemistry schemes or mixed solution phase and solid phase schemes. Examples of synthetic routes are described in the examples below. Unless otherwise indicated, the compounds of formula (I) and related formulae obtained as racemic mixtures may be separated to provide enantiomerically enriched mixtures or pure enantiomers.

Commercially available starting materials used in the following experimental descriptions were purchased from Aldrich or Fluka, unless otherwise indicated.

The HPLC, NMR and MS data provided in the examples below were obtained as follows:

HPLC data:

the method A comprises the following steps: HPLC column: xbridge C8 column 50 mm x 4.6 mm, flow rate is 2 mL/min; from 0.1% TFA in H₂Gradient 8 min from solution in O to 0.07% TFA in ACN.

The method B comprises the following steps: HPLC column: CHIRALCEL OJ-H, 250X 4.6 mm, 5 μm, 30 ℃. Gradient: Hexane/ISOH 80/200.1% TEA at a flow rate of 1 mL/min.

The method C comprises the following steps: HPLC column: hichrom, Kromasil Eternity, 2.5. mu. m C18, 150X 4.6 mm, flow rate 1 mL/min; from 95:5 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) to 5:95 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) gradient at 6.0 min, then at 5:95 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) was held for 4.6 minutes.

The method D comprises the following steps: HPLC column: phenomenex Luna 5 mu m C18 (2), 100X 4.6 mm (with protective cylinder), and the flow rate is 2 mL/min; from 95:5 ([0.1% (V/V) formic acid in H₂Solution in O]Solutions of [0.1% (V/V) formic acid in MeCN]) To 5:95 ([0.1% (V/V) formic acid in H₂Solution in O]Solutions of [0.1% (V/V) formic acid in MeCN]) Gradient of 3.5 min, then formic acid in H at 5:95 ([0.1% (V/V))₂Solution in O]Solutions of [0.1% (V/V) formic acid in MeCN ]) Hold for 2 minutes.

The method E comprises the following steps: HPLC column: waters Sunfire 5. mu. m C18, 150X 4.6mm (with protective cartridge) at a flow rate of 1 mL/min; from 95:5 ([0.1% (V/V) formic acid in H₂Solution in O]0.1% (V/V) formic acid in MeOH]) A30 min gradient to a 0.1% (V/V) formic acid solution in MeOH, then a 0.1% (V/V) formic acid solution in MeOH for 5 minutes.

Method F: HPLC column: phenomenex, Gemini NX, 3 mu m C18, 100X 4.6mm, and the flow rate is 2 mL/min; from 95:5 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) to 5:95 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) gradient at 3.5 min, then at 5:95 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) was held for 1.5 minutes.

Method G: HPLC column: waters Xterra MS 5. mu. m C18, 100X 4.6mm (with protective cartridge) at a flow rate of 2 mL/min; from 95:5 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) to 5:95 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) gradient at 3.5 min, then at 5:95 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeCN) was held for 1.5 minutes.

Method H: apply the gradient of method H to the HPLC column: supelco, Ascentis ^ Express C18 or Hichrom Halo C18, 2.7 μm C18, 100X 4.6 mm.

Method of producing a composite materialI: HPLC column: waters Xbridge 5. mu. m C18, 150X 4.6 mm (with protective cartridge) at a flow rate of 1 mL/min; from 95:5 ([10 mM ammonium bicarbonate in H)₂Solution in O]MeOH) to a 22 min gradient of MeOH, then held in the MeOH for 4 min.

Method J: HPLC column: chiralpak IC, 5 μm, 250X 4.6 mm (chiral technologies), flow rate 1 mL/min; isocratic hexane-EtOH-TFA-DEA 70: 30: 0.1: 0.1.

Method K: HPLC column: chiralpak OJH, 5 μm, 250 × 4.6 mm (chiral technologies), flow rate 1 mL/min; isocratic hexane-EtOH-TFA 97: 3: 0.1.

The method L comprises the following steps: HPLC column: chiralpak AD-RH, 5 μm, 150X 4.6 mm (chiral technologies), flow rate 1 mL/min, at 40 ℃; isocratic buffer pH 2, 0.1M KPF₆.- ACN 65: 35。

Method M: HPLC column: chiralpak OD-RH, 5 μm, 150X 4.6 mm (chiral technologies), flow rate 1 mL/min; isocratic buffer pH 2, 0.1M HClO₄/NaClO₄.- ACN 60: 30。

Ultraviolet detection (maxplot) for all methods.

Mass spectrometry：

The method A comprises the following steps: LC/MS Waters ZMD (ESI); GC/MS: GC Agilent 6890N and MS Agilent 5973.

The method B comprises the following steps: UPLC/MS: waters Acquity, column Waters Acquity UPLC BEH C181.7 m 2.1 x 50 mm, conditions: solvent A (10 mM ammonium acetate in water + 5% ACN), solvent B (ACN), gradient 5% B to 100% B (over 3 min), UV detection (PDA, 230-.

¹ H-NMR Data of：

Bruker DPX-300 MHz, unless otherwise noted.

Preparation type HPLC And (3) purification:

preparative HPLC purifications were performed with the HPLC waters Prep LC 4000 system (equipped with columns of PrepMS C1810 m, 50X 300 mm) unless otherwise indicated. With ACN/H₂O or ACN/H₂An O/TFA (0.1%) gradient was used for all HPLC purifications.

The masses being guided Autoprep (MD Autoprep) And (3) purification:

preparative HPLC purification was performed with mass-directed, automated purification Fractionlynx from Waters equipped with a Sunfire Prep C18 OBD column 19X 100 mm 5 m, unless otherwise indicated. With ACN/H₂O or ACN/H₂An O/HCOOH (0.1%) gradient was used for all HPLC purifications.

In a single mode microwave reactor Emrys^TMOptimiers or initiators^TMMicrowave chemistry was performed on Sixty (from Biotage) or Explorer (from CEM).

General operation:

general operation 1: general procedure for Aminoxime moiety formation

To a solution of the nitrile derivative (1 eq) in EtOH (1-5 mL/mmol of nitrile derivative) was added 50% NH₂Aqueous OH (5 equivalents). The resulting mixture is stirred at a temperature of from room temperature to 80 ℃ for 1 to 72 hours. In the case where the desired compound precipitated, the precipitate was filtered off, washed with a sufficient solvent (such as EtOH, iPrOH or water), and then dried under reduced pressure to give the desired ammoxim derivative. In all other cases, the reaction mixture was concentrated under reduced pressure, diluted with sufficient solvent (such as water or iPrOH) until precipitation. The precipitate is filtered off and washed with sufficient solvent (such as iPrOH or water) and then dried under reduced pressure to give the desired ammoxim derivative. When no precipitation occurred, the concentrated mixture was dissolved in EtOAc and water In (1), the organic layer was washed with water (2 times) and brine (2 times), then over MgSO₄Drying, filtration and concentration under vacuum gave the desired ammoxine derivative.

General operation 2: general procedure for Aminoxime moiety formation

To a solution of the nitrile derivative (1 eq) in EtOH (1-5 mL/mmol of nitrile derivative) was added NH₂OH.HCl (1.1 equiv.) and Et₃N (1.2 equiv.). The resulting mixture is stirred at a temperature of from room temperature to 80 ℃ for 1 to 72 hours. In the case where the desired compound precipitated, the precipitate was filtered off, washed with a sufficient solvent (such as EtOH, iPrOH or water), and then dried under reduced pressure to give the desired ammoxim derivative. In all other cases, the reaction mixture was concentrated under reduced pressure, diluted with sufficient solvent (such as water or iPrOH) until precipitation. The precipitate is filtered off and washed with sufficient solvent (such as iPrOH or water) and then dried under reduced pressure to give the desired ammoxim derivative. When no precipitation occurred, the concentrated mixture was dissolved in EtOAc and water, the organic layer was washed with water (2 times) and brine (2 times), then over MgSO₄Drying, filtration and concentration under vacuum gave the desired ammoxine derivative.

General operation 3: general procedure for formation of oxadiazole Ring

DIEA (2.0-2.2 equiv.) and HATU (1.0-1.1 equiv.) were added to a solution of the carboxylic acid derivative (1 equiv.) in anhydrous DMF (4 mL/mmol of carboxylic acid derivative) cooled at 0 ℃. The resulting mixture was stirred at 0 ℃ for a period of 5-30 minutes. Then, the pure ammoxine derivative (1.0-1.2 equivalents) was added or added as a DMF solution (2 mL/mmol of the ammoxine derivative). The resulting mixture was stirred at 0 ℃ or RT for a period of 30 minutes to 18 hours. With sufficient solvent (such as Et)₂O, MTBE or EtOAc) and then washed with water and brine. The aqueous layer was extracted 1 time. The combined organic layers were dried (MgSO)₄Or Na₂SO₄) And the solvent was removed under reduced pressure.The residue was dissolved with toluene (6 mL/mmol carboxylic acid derivative) and pyridine (2 mL/mmol carboxylic acid derivative) or with ACN (8.5 mL/mmol carboxylic acid derivative). The resulting mixture was heated at a temperature between 80 ℃ and reflux for a period of 12-72 hours. With sufficient solvent (such as Et)₂O, MTBE or EtOAc) and then washed with water and brine. The aqueous layer was extracted 1 time. The combined organic layers were dried (MgSO)₄Or Na₂SO₄) And the solvent was evaporated under reduced pressure. Purification by flash chromatography or precipitation gives the desired oxadiazole derivative.

General operation 4: general procedure for formation of oxadiazole Ring

DIEA (2.0-2.2 equiv.) and HATU (1.0-1.1 equiv.) were added to a solution of the carboxylic acid derivative (1 equiv.) in anhydrous DMF (4 mL/mmol of carboxylic acid derivative) cooled at 0 ℃. The resulting mixture was stirred at 0 ℃ for a period of 5-30 minutes. Then, the pure ammoxine derivative (1.0-1.2 equivalents) was added or added as a DMF solution (2 mL/mmol of the ammoxine derivative). The resulting mixture was stirred at 0 ℃ or RT for a period of 30 minutes to 18 hours. With sufficient solvent (such as Et)₂O, MTBE or EtOAc) and then washed with water and brine. The aqueous layer was extracted 1 time. The combined organic layers were dried (MgSO)₄Or Na₂SO₄) And the solvent was removed under reduced pressure. The residue was dissolved with ACN (8.5 mL/mmol of carboxylic acid derivative). The resulting mixture was heated at 150 ℃ for 30 min under Microwave (MW) irradiation. With sufficient solvent (such as Et)₂O, MTBE or EtOAc) and then washed with water and brine. The aqueous layer was extracted 1 time. The combined organic layers were dried (MgSO)₄Or Na₂SO₄) And the solvent was evaporated under reduced pressure. Purification by flash chromatography or precipitation gives the desired oxadiazole derivative.

General operation 5: general procedure for formation of oxadiazole Ring

In a microwave vessel, add the carboxylic acid derivative (1 equivalent)) Aminoxime derivative (1.1 equiv.), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.4 equiv.) and CH₃CN (1 mL/mmol carboxylic acid). The reaction mixture was stirred at room temperature for 2-18 hours. Pyridine (0.3 mL/mmol carboxylic acid) was then added and the mixture was heated under microwave irradiation at 150 ℃ for 30 minutes. The solvent was removed in vacuo, in DCM and H₂The residue was partitioned between O. Separating the layers with H₂The organic layer was washed and the mixture was passed through a hydrophobic frit filter plate (frat). The solvent is removed in vacuo and the residue is purified by recrystallization, precipitation or flash chromatography to give the desired oxadiazole derivative.

General operation 6 : benzyl alcohol oxidation

Benzyl alcohol derivative (1 eq) was dissolved in dioxane (7 mL/mmol benzyl alcohol) and manganese dioxide (8 eq) was added. The mixture was heated at 70 ℃ overnight and the solvent was removed in vacuo. The residue was triturated with a mixture of petrol/ether to give the corresponding benzaldehyde derivative.

General operation 7 : reductive amination using amino acid derivatives

Sodium cyanoborohydride (1.1 equiv.) was added to a solution of the aldehyde derivative (1 equiv.; 0.25 mmol) and the appropriate amino acid (2 equiv.) in a 1:1 mixture of methanol/DCM (24 mL/mmol) and acetic acid (2.5 equiv.). The mixture was stirred at room temperature overnight and filtered through a glass frit filter plate under positive pressure. To the filtrate was added formaldehyde (37% aqueous solution; 10 equivalents) followed by AcOH until the pH was in the range of 3-4. To the resulting mixture was added sodium cyanoborohydride (5 equivalents), the mixture was stirred for 16 hours, the solvent was removed in vacuo, and the residue was purified by reverse phase HPLC.

General operation 8 ：Hydrolysis of tert-butyl ester

To the tert-butyl ester derivative (1 equivalent) is addedHydrochloric acid in dioxane (4N, 20-50 equivalents), and the reaction mixture was stirred at room temperature for 1 to 24 hours. The solution is then evaporated to dryness by evaporation from a solvent (such as CH)₃CN, DCM, MTBE or Et₂O) to yield the title compound. When no precipitation occurred, the title compound was purified by reverse phase HPLC or mass directed Autoprep.

General operation 9 ：Hydrolysis of methyl or ethyl ester

To a solution of methyl or ethyl ester derivative (1 equivalent) in MeOH or EtOH (5 mL/mmol ester derivative) or MeOH/THF 1:1 or EtOH/THF 1:1 (5 mL/mmol ester derivative) was added sodium hydroxide (5M, 5 equivalents) and the reaction mixture was stirred at room temperature for 1 to 24 hours. The solution was evaporated to dryness. The residue was dissolved with EtOAc and washed with 1N aqueous HCl and brine. Drying (MgSO)₄) The organic layer was concentrated in vacuo to afford the title compound.

General operation 10: substitution of benzyl bromide with primary and secondary amines

To benzyl bromide derivative (1 equivalent) in CH₃CN or DMF (1.5-3 mL/mmol benzyl bromide derivative), adding K₂CO₃Or NaHCO₃(2-3 equivalents) and a primary or secondary amine (1.2 equivalents). The reaction mixture was stirred at room temperature or at 60 ℃ overnight. The solvent was removed in vacuo, the resulting mixture was diluted with water, extracted with EtOAc, washed with saturated NaCl solution, over MgSO ₄Dried and concentrated under vacuum. Purification by flash chromatography or recrystallization gives the desired benzylamine derivative.

General operation 11 ：Substitution of benzylamines

To benzylamine derivative (1 equivalent) in CH₃CN or DMF (1.5-3 mL/mmol benzylamine derivative), K is added₂CO₃Or NaHCO₃(2-3 equivalents) and an electrophile such as bromoethylAcid alkyl ester derivative (1.2 equivalents). The reaction mixture was stirred at room temperature or at 60 ℃ overnight. The solvent was removed in vacuo and the resulting mixture was diluted with water, extracted with EtOAc, washed with saturated NaCl solution and MgSO₄Dried and concentrated under vacuum. Purification by flash chromatography or recrystallization gives the desired benzylamine derivative.

General operation 12 ：Preparation of N-oxide (N-oxide)

The compound of formula (I) (1 equivalent) was dissolved in DCM (10 mL/mmol oxadiazole derivative). 3-Chloroperoxybenzoic acid (1 eq.) was added at 0 ℃. The mixture was stirred at room temperature for 1 hour. This was then evaporated and the crude product was purified on MD autoprep to give the desired N-oxide derivative.

Intermediates 1: 2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Formic acid

Step (ii) of 1: 4- Bromine compound -3-( Trifluoromethyl radical ) Benzoic acid methyl ester

To a suspension of 4-bromo-3- (trifluoromethyl) benzoic acid (Acceledev 000625, 15 g; 56 mmol) in MeOH (300 mL) at room temperature was added thionyl chloride (16 mL; 223 mmol) dropwise over 15 min. The reaction mixture was stirred at room temperature for 12 hours. The solvent was concentrated and the crude residue was diluted with EtOAc (500 mL). With saturated NaHCO ₃The organic layer was washed with aqueous solution (200 mL), water (200 mL), brine (200 mL), and MgSO₄Dried and concentrated to give the title compound as an orange solid (14.8 g, 94%).¹H NMR (DMSO-d₆300 MHz) delta 8.26 (m, 1H), 8.14-8.13 (m, 2H), 3.93 (s, 3H). HPLC (method A) Rt 4.71 min (purity: 99.0%).

Step (ii) of 2: 2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Formic acid methyl ester

In N₂Methyl 4-bromo-3- (trifluoromethyl) benzoate (6 g; 21 mmol), o-tolylboronic acid (3.2 g; 23 mmol), potassium carbonate (14.7 g; 106 mmol), tetrakis (triphenylphosphine) palladium (0) (2.5 g; 2.12 mmol) were dissolved in toluene (30 mL) and water (30 mL) under an atmosphere. The reaction mixture was purged with vacuum for 5 minutes, then with N₂Degassed and refluxed for 3 hours. The reaction mixture was cooled to room temperature, filtered over a pad of celite, and washed with toluene (200 mL). The filtrate was concentrated to give a brown oil, which was dissolved in EtOAc (200 mL). With saturated NaHCO₃The organic layer was washed with aqueous solution (50 mL), water (50 mL) and brine (50 mL), over MgSO₄Dried and concentrated to give the title compound as a brown oil (6.4 g, quantitative). HPLC (method A) Rt 5.33 min (purity: 60.0%).

Step (ii) of 3: 2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Formic acid

A solution of methyl 2' -methyl-2- (trifluoromethyl) biphenyl-4-carboxylate (5 g; 17 mmol) in EtOH (150 mL) was treated with sodium hydroxide (10.2 mL; 5M; 51 mmol) at room temperature. The reaction mixture was stirred at 60 ℃ for 2 hours. The reaction mixture was concentrated to give a brown solid, which was dissolved in water (300 mL) and the aqueous phase was washed 2 times with EtOAc. The aqueous phase was acidified with HCl cc to pH 2 and then concentrated to precipitate (half volume). The suspension was filtered to give the title compound as a light brown solid.¹H NMR (DMSO-d₆, 300 MHz) δ13.55 (br s, 1H), 8.31 (s, 1H), 826-8.23 (d, J =7.9 Hz, 1H), 7.51-7.48 (d, J =7.9 Hz 1H), 7.37-7.12 (m, 4H), 1.99 (s, 3H). LC/MS (method A): 278.9 (M-H)^-. HPLC (method A) Rt 4.57 min (purity: 98.7%).

Intermediates 2: 2'-( Difluoromethyl group )-2- Methyl biphenyl -4- Formic acid

Step (ii) of 1: 2'-( Difluoromethyl group )-2- Methyl biphenyl -4- Formic acid methyl ester

In N₂Preparation of methyl 3-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzoate (Combiblocks PN-8756, 1.9 g, 6.7 mmol), 1-bromo-2-difluoromethyl-benzene (Fluorochem 023878, 1.7 g, 8.1 mmol), cesium fluoride (3.1 g, 20 mmol) and PdCl under atmosphere₂(PPh₃)₂(142 mg, 0.20 mmol) in dioxane (20 mL) and water (10 mL). The resulting mixture was heated at 90 ℃ for 2 hours. The reaction mixture was diluted with MTBE (60 mL) and washed with water (2 × 50 mL) and brine (50 mL). The aqueous layer was extracted with MTBE (50 mL). The combined organic layers were dried (MgSO) ₄) And concentrated under reduced pressure. After purification by flash chromatography (silica gel, DCM/heptane) the title compound was obtained as a colourless oil (1.4 g, 76%). HPLC (method A) Rt 5.0 min (purity: 99.1%).¹H NMR (DMSO-d₆, 300 MHz) δ7.93 (s, 1H), 7.84 (dd, J = 7.9, 1.4 Hz, 1H), 7.75 (m, 1H), 7.67-7.56 (m, 2H), 7.28 (m, 2H), 6.55 (t, J = 55 Hz, 1H), 3.88 (s, 3H), 2.06 (s, 3H)。

Step (ii) of 2: 2'-( Difluoromethyl group )-2- Methyl biphenyl -4- Formic acid

Aqueous 5N NaOH (1.5 mL, 7.5 mmol) was added to a solution of methyl 2' - (difluoromethyl) -2-methylbiphenyl-4-carboxylate (1.4 g, 5.1 mmol) in EtOH (15 mL). The resulting mixture was heated at 70 ℃ for 1 hour and then evaporated. The residue was dissolved with water (25 mL) and 5N aqueous HCl (3 mL) and extracted with MTBE (2X50 mL). The organic layers were washed with brine (25 mL), combined and dried (MgSO)₄) And concentrated under reduced pressure to give the title compound as a white powder (1.3 g, 93%). HPLC (method A) Rt 4.2 min (purity: 98.9%). LC/MS (method B) 261.1 (M-H)^-。¹H NMR (DMSO-d₆, 300 MHz) δ13.1 (s, 1H), 7.90 (s, 1H), 7.82 (dd, J = 7.9, 1.4 Hz, 1H), 7.74 (m, 1H), 7.66-7.55 (m, 2H), 7.26 (m, 2H), 6.55 (t, J = 55 Hz, 1H), 2.05 (s, 3H)。

Intermediates 3: 2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Formic acid

Step (ii) of 1: 4- Bromine compound -3-( Bromomethyl group ) Benzoic acid methyl ester

In N₂To methyl 4-bromo-3-methylbenzoate (Aldrich 532878, 50 g; 218 mmol) in CHCl under an atmosphere₃NBS (47 g; 262 mmol) and. alpha.,. alpha.' -azoisobutyronitrile (720 mg; 4.4 mmol) were added to the solution in (1L) in one portion. The mixture was stirred at 70 ℃ for 2 days. The reaction mixture was cooled to room temperature and water (500 mL) was added. With 50 mL NaHCO ₃The organic layer was washed with saturated solution, water (340 mL), then brine (500 mL), MgSO₄Dried and concentrated to give the title compound asYellow solid. It was washed with pentane (2 × 500 mL) to give the title compound as a yellow solid.¹H NMR (DMSO-d₆300 MHz) δ 8.24 (d, J = 1.9 Hz, 1H), 7.88-7.82 (m, 2H), 4.87 (s, 2H), 3.91 (s, 3H). HPLC (method A) Rt 4.44 min (purity: 97.9%).

Step (ii) of 2: 4- Bromine compound -3-( Methoxymethyl group ) Benzoic acid methyl ester

A solution of methyl 4-bromo-3- (bromomethyl) benzoate (38 g; 122 mmol) in MeOH (1.1L) was refluxed for 4 days. After concentration, the mixture was partitioned between EtOAc (500 mL) and water (200 mL). With 5% NaHCO₃The organic layer was washed with aqueous solution (200 mL), brine (200 mL), and MgSO₄Dried and concentrated to give the title compound as a beige solid (30 g, 94%).¹H NMR (DMSO-d₆300 MHz) delta 8.06-8.05 (m, 1H), 7.83 (m, 2H), 4.54 (m, 2H), 3.90 (s, 3H), 3.45 (s, 3H). LC/MS (method B) 227.2 (M-H)^-. HPLC (method A) Rt 4.42 min (purity: 93.0%).

Step (ii) of 3: 2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Formic acid methyl ester

In N₂Methyl 4-bromo-3- (methoxymethyl) benzoate (40 g; 154 mmol), o-tolylboronic acid (23 g; 170 mmol) and K were reacted under an atmosphere ₂CO₃(107 g; 772 mmol), tetrakis (triphenylphosphine) palladium (0) (1.8 g; 1.5 mmol) were dissolved in toluene (200 mL) and water (200 mL). The reaction mixture was purged with vacuum and then with N₂Degassed and refluxed for another 1 hour. The reaction mixture was cooled to room temperature, filtered over a pad of celite, and washed with EtOAc (1L). The filtrate was concentrated to give a yellow oil, which was dissolved in EtOAc (800 mL). With saturated NaHCO₃The organic layer was washed with aqueous (250 mL), water (250 mL) and brine (250 mL) over MgSO₄Drying, and concentrating the mixture,the title compound was obtained as a yellow oil which was used without further purification (42 g, quantitative). HPLC (method A) Rt 5.34 min (purity: 89.4%).

Step (ii) of 4: 2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Formic acid

A solution of methyl 2- (methoxymethyl) -2' -methylbiphenyl-4-carboxylate (40 g; 148 mmol; 1 eq) in EtOH (1.2L) was treated with NaOH (89 mL; 5M; 445 mmol) at room temperature. The reaction mixture was stirred at 60 ℃ for 1 hour. The reaction mixture was cooled to room temperature and concentrated to give a yellow solid, which was dissolved in water (800 mL) and the aqueous phase was washed 2 times with EtOAc. The aqueous phase was acidified with HCl cc (40 mL) to pH 2 and extracted with EtOAc (2X400 mL). The combined organic phases were washed with brine, MgSO ₄Dried and concentrated to give the title compound as a yellow solid (35 g, 92%).¹H NMR (DMSO-d₆300 MHz) delta 12.99 (br s, 1H), 8.09 (s, 1H), 7.92-7.89 (m, 1H), 7.33-7.22 (m, 4H), 7.10-7.08 (m, 1H), 4.11 (m, 2H), 3.18 (s, 3H), 1.99 (s, 3H). LC/MS (method B): 255.2 (M-H)^-. HPLC (method A) Rt 4.52 min (purity: 96.4%).

Intermediates 4: 2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Formic acid

In N₂A mixture of methyl 4-bromo-3-methylbenzoate (20 g, 87 mmol), 2- (trifluoromethyl) phenylboronic acid (24.9 g, 131 mmol), potassium carbonate (24 g, 175 mmol) and bis (tricyclohexylphosphine) palladium (II) dichloride (65 mg, 0.1 mmol) in dioxane (200 mL) and water (50 mL) was prepared under an atmosphere. Mixing the mixtureHeating at 100 deg.C for 3 hr. Aqueous 5N NaOH (100 mL) was added and the reaction mixture was stirred at 100 ℃ for an additional 1 hour. The reaction mixture was cooled at room temperature, and the aqueous layer was removed. The organic layer was filtered through a celite pad, concentrated to 75 mL under reduced pressure, diluted with water (125 mL), and washed with MTBE (2 × 200 mL). The aqueous layer was acidified to pH 1 with 5N aqueous HCl (25 mL) and extracted with MTBE (2X100 mL). The organic layers were combined and dried (Na)₂SO₄) And filtered through a cilat pad. The solution was concentrated to 100 mL and heptane (200 mL) was added. The mixture was concentrated to 100 mL. The precipitate was filtered off and washed 2 times with heptane, then dried under reduced pressure to give the title compound as a white powder (22.5 g, 92%). HPLC (method A) Rt 4.4 min (purity: 100%). LC/MS (method B) 279.0 (M-H) ^-。¹H NMR (DMSO-d₆, 300 MHz) δ13.0 (s, 1H), 7.87 (m, 2H), 7.80 (dd, J = 7.9, 1.6 Hz, 1H), 7.75 (m, 1H), 7.64 (m, 1H), 7.34 (d, J = 7.6 Hz, 1H), 7.23 (d, J = 7.9 Hz, 1H), 2.02 (s, 3H)。

Intermediates 5 ： 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ] Tert-butyl butyrate

Step (ii) of 1: 3- Fluorine -4-[( Methylamino radical ) Methyl radical ] Benzonitrile

A solution of 4-cyano-2-fluoro-benzyl bromide (15 g, 0.07 mol) in dry THF (50 mL) was added to 40% aqueous methylamine solution (400 mL) at 5-10 deg.C with stirringStirring for 5 min. The mixture was extracted with DCM and the organic layer was washed with Na₂SO₄Dried and concentrated in vacuo to give the title compound as a brown liquid (10 g, 89%).¹H NMR (DMSO-d₆, 400 MHz) δ7.77 (m, 1H), 7.68-7.61 (m, 2H), 3.71 (s, 2H), 2.24 (s, 3H)。

Step (ii) of 2: 2-[(2- Fluorine -4- Isocyanobenzyl radical )( Methyl radical ) Amino group ] Tert-butyl butyrate

From 3-fluoro-4- [ (methylamino) methyl according to general procedure 11]The title compound was prepared starting from benzonitrile and tert-butyl 2-bromobutyrate. It was isolated as a colorless liquid (9.5 g, 71%).¹H NMR (DMSO-d₆, 400 MHz) δ7.80 (m, 1H), 7.68 (m, 1H), 7.61 (t, J =7.4 Hz, 1H), 3.85-3.70 (dd, J = 14.9, 32.1 Hz, 2H), 3.08-3.04 (t, J = 7.6 Hz, 1H), 2.19 (3H, s), 1.70-1.63 (m, 2H), 1.42 (s, 9H), 0.87-0.83 (t, J = 7.3 Hz, 3H)。

Step (ii) of 3: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ] Tert-butyl butyrate

From 2- [ (2-fluoro-4-isocyanobenzyl) (methyl) amino]The title compound was prepared starting from tert-butyl butyrate. It was isolated as a white solid.¹H NMR (DMSO-d₆400 MHz) δ 9.72 (s, 1H), 7.49 (m, 1H), 7.39 (m, 2H), 5.85 (s, 2H), 3.77-3.62 (m, 2H), 3.04 (t, J =7.5 Hz, 1H), 2.19 (s, 3H), 1.61 (m, 2H), 1.44 (s, 9H), 0.85 (t, J =7.3 Hz, 3H). LC/MS (method A) 307.3 (M + H)⁺. HPLC (method A) Rt 2.3 min (purity: 98.2%).

Intermediates 6: N-((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-D- Alanine methyl ester

Step (ii) of 1: N-[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ]-D- Alanine methyl ester and N-[(1R)-1-(4- cyanophenyl group ) Ethyl radical ]-L- Alanine methyl ester

To 4- [ (1R) 1-aminoethyl group in DMF (34 mL)]To benzonitrile (Haoyuran Chemex press, 3.4 g; 23 mmol) were added sodium bicarbonate (3.9 g; 46 mmol) and methyl 2-bromopropionate (2.8 mL; 25 mmol). The reaction mixture was stirred at 75 ℃ overnight. EtOAc is added to the reaction mixture and the organic phase is washed with H₂O Wash 3 times and brine 1 time, then MgSO₄The organic layer was dried, filtered, and concentrated. The crude product was purified by flash chromatography (EtOAc/heptane gradient from 5:95 to 25: 75) to yield N- [ (1R) -1- (4-cyanophenyl) ethyl]-D-alanine methyl ester (first eluting compound) white solid (2.1 g, 40%) and N- [ (1R) -1- (4-cyanophenyl) ethyl]L-alanine methyl ester (second eluting compound) orange solid (2.3 g, 43%).

N-[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ]-D- Alanine methyl ester : ¹H NMR (DMSO-d₆, 300 MHz) δ 7.78 (d, J = 8.4 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 3.75 (q, J = 6.3 Hz, 1H), 3.62 (s, 3H), 2.95-2.81 (m, 1H), 2.71-2.58 (m, 1H), 1.22 (d,J = 6.4 Hz, 3H), 1.14 (d, J = 7.0 Hz, 3H). LC/MS (method B): 233.0 (M + H)⁺. HPLC (method A) Rt 1.58 min (purity: 70.7%).

N-[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ]-L- Alanine methyl ester : ¹H NMR (DMSO-d₆300 MHz) δ 7.77 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.1 Hz, 2H), 3.86 (q, J = 6.1 Hz, 1H), 3.46 (s, 3H), 3.26-3.13 (m, 1H), 2.53 (m, 1H), 1.23 (d, J = 6.6 Hz, 3H), 1.17 (d, J = 6.8 Hz, 3H). LC/MS (method B) 233.0 (M + H), n.d (M-H). HPLC (method A) Rt 1.49 min (purity: 72.4%).

Step (ii) of 2: N-((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-D- Alanine methyl ester

Following general procedure 1, starting from N- [ (1R) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from-D-alanine methyl ester. It was isolated as a slightly yellow oil (2.3 g, 93%).¹H NMR (DMSO-d₆300 MHz) δ 9.56 (s, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.28 (d, J = 8.3 Hz, 2H), 5.76 (s, 2H), 3.72-3.57 (m, 4H), 3.03-2.82 (m, 1H), 2.49-2.41 (m, 1H), 1.21 (d, J = 6.6 Hz, 3H), 1.13 (d, J = 7.1 Hz, 3H). LC/MS (method B) 266.1 (M + H)⁺。

Intermediates 7: N-[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ]-L- Alanine methyl ester

Following general procedure 1, starting from N- [ (1R) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from-L-alanine methyl ester (for its synthesis see intermediate 6, step 1). It was isolated as a yellow oil (2.1 g, 80%).¹H NMR (DMSO-d₆300 MHz) δ 9.57 (s, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.30 (d, J = 8.3 Hz, 2H), 5.76 (s, 2H), 3.82-3.70 (m, 1H), 3.49 (s, 3H), 3.26-3.13 (m, 1H), 2.35-2.26 (m, 1H), 1.23 (d, J = 6.6 Hz, 3H), 1.17 (d, J = 6.8 Hz, 3H). LC/MS (method A) 266.1 (M + H)⁺。

Intermediates 8: N-((1S)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

Step (ii) of 1: N-[(1S)-1-(4- Cyanophenyl group ) Ethyl radical ] Glycine tert-butyl ester

Following general procedure 11, starting from 4- [ (1S) -1-aminoethyl)]Benzonitrile (haoyuran Chemexpress) and tert-butyl bromoacetate. It was isolated as a yellow oil (1.3g, 73%).¹H NMR (DMSO-d₆300 MHz) δ 7.78 (d, J = 8.3 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 3.81 (m, 1H), 3.00 (m, 2H), 2.45 (m, 1H), 1.37 (s, 9H), 1.24 (d, J = 6.6 Hz, 3H). LC/MS (method B): 233.0 (M + H)⁺。

Step (ii) of 2: N-((1S)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

Following general procedure 1, starting from N- [ (1S) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from tert-butyl glycinate and isolated as a white foam (1.3g, 87%).¹H NMR (DMSO-d₆, 300 MHz) δ 9.57 (s, 1H), 7.60 (d, J = 8.3 Hz, 2H), 7.28 (d, J = 8.3 Hz, 2H), 5.76 (brs, 2H), 3.73 (m, 1H), 3.43 (m, 1H), 3.01 (m, 2H), 1.38 (s, 9H), 1.24 (d, J = 6.6 Hz, 3H)。

Intermediates 9: N-((1R)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

Step (ii) of 1: N-[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ] Glycine tert-butyl ester

Following general procedure 11, starting from 4- [ (1R) -1-aminoethyl)]Benzonitrile (haoyuran Chemexpress) and tert-butyl bromoacetate the title compound was prepared starting and isolated as a yellow oil.¹H NMR (DMSO-d₆300 MHz) δ 7.78 (d, J = 8.2 Hz, 2H), 7.51 (d, J = 8.2 Hz, 2H), 3.82 (m, 1H), 3.00 (m, 2H), 2.47 (m, 1H), 1.37 (s, 9H), 1.24 (d, J = 6.6 Hz, 3H). HPLC (method A) Rt 2.77 min (purity: 99.4%).

Step (ii) of 2: N-((1R)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

Following general procedure 1, starting from N- [ (1R) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from tert-butyl glycinate and isolated as a yellow oil (800 mg, 86%).¹H NMR (DMSO-d₆, 300 MHz) δ 9.57 (s, 1H), 7.60 (d, J = 8.2 Hz, 2H), 7.28 (d, J = 8.2 Hz, 2H), 5.76 (brs, 2H), 3.73 (m, 1H), 3.43 (m, 1H), 3.01 (m, 2H), 1.38 (s, 9H), 1.24 (d, J = 6.6 Hz, 3H)。

Intermediates 10: N-((1S)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-D- Alanine methyl ester

Step (ii) of 1: N-[(1S)-1-(4- Cyanophenyl group ) Ethyl radical ]-D- Alanine methyl ester and N-[(1S)-1-(4- cyanophenyl group ) Ethyl radical ]-L- Alanine methyl ester

To 4- [ (1S) -1-aminoethyl group in DMF (15 mL)]To benzonitrile (Haoyuran Chemex press, 1 g; 6.8 mmol) were added sodium bicarbonate (1.2 g; 14 mmol) and methyl 2-bromopropionate (0.84 mL; 7.5 mmol). The reaction mixture was stirred at 75 ℃ overnight. EtOAc is added and the organic phase is washed with H₂O Wash 3 times and brine 1 time, then MgSO₄The organic layer was dried, filtered, and concentrated. The crude product was purified by flash chromatography (EtOAc/heptane gradient from 5:95 to 25: 75) to yield N- [ (1S) -1- (4-cyanophenyl) ethyl]-L-alanine methyl ester (first wash)Des compound) was a pale yellow solid (480 mg; 30%) and N- [ (1S) -1- (4-cyanophenyl) ethyl]methyl-D-alanine (second eluting compound) as a yellow solid (470 mg; 30%).

N-[(1S)-1-(4- Cyanophenyl group ) Ethyl radical ]-L- Alanine methyl ester : ¹H NMR (DMSO-d₆300 MHz) δ 7.77 (d, J = 8.4 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 3.74 (m, 1H), 3.61 (s, 3H), 2.93-2.81 (m, 1H), 2.64 (m, 1H), 1.21 (d, J = 6.5 Hz, 3H), 1.13 (d, J = 7.0 Hz, 3H). LC/MS (method B): 233.1 (M + H)⁺. HPLC (method A) Rt 1.57 min (purity: 79%).

N-[(1S)-1-(4- Cyanophenyl group ) Ethyl radical ]-D- Alanine methyl ester : 1H NMR (DMSO-d₆300 MHz) δ 7.77 (d, J = 8.3 Hz, 2H), 7.53 (d, J = 8.3 Hz, 2H), 3.85 (q, J = 6.1 Hz, 1H), 3.45 (s, 3H), 3.25-3.13 (m, 1H), 2.53 (m, 1H), 1.23 (d, J = 6.6 Hz, 3H), 1.16 (d, J = 6.8 Hz, 3H). LC/MS (method B): 233.0 (M + H)⁺. HPLC (method A) Rt 1.52 min (purity: 89.2%).

Step (ii) of 2: N-((1S)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-D- Alanine methyl ester

Following general procedure 1, starting from N- [ (1S) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from-D-alanine methyl ester. It was isolated as a colorless oil (500 mg, 93%).¹H NMR (DMSO-d₆, 300 MHz) δ 9.57 (s, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H), 5.76 (brs, 2H), 3.76 (m, 1H), 3.49 (s, 3H), 3.27-3.12 (m, 1H), 2.31 (m, 1H), 1.23 (d, J = 6.6 Hz, 3H), 1.17 (d, J = 6.8 Hz, 3H) In that respect LC/MS (method B) 266.1 (M + H)⁺。

Intermediates 11: N-((1S)-1-{4-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-L- Alanine methyl ester

Following general procedure 1, starting from N- [ (1S) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from-L-alanine methyl ester (for its synthesis see intermediate 10, step 1). It was isolated as a colorless oil. ¹H NMR (DMSO-d₆300 MHz) δ 9.56 (s, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.27 (d, J = 8.3 Hz, 2H), 5.75 (brs, 2H), 3.66 (m,1H), 3.61(s, 3H), 2.92 (m,1H), 2.49-2.41 (m,1H), 1.21 (d, J = 6.6 Hz, 3H), 1.13 (d, J = 6.9 Hz, 3H). LC/MS (method A) 266.1 (M + H)⁺。

Intermediates 12: (2S)-2-[((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Amino group ] Butyric acid ethyl ester

Step (ii) of 1: (2S)-2-{[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ] Amino group } Butyric acid ethyl ester and (2R)-2-{[(1R)-1-(4- cyanophenyl group ) Ethyl radical ] Amino group } Butyric acid ethyl ester

The title compound was prepared following general procedure 11 starting from 4- [ (1R) -1-aminoethyl) ] benzonitrile (Haoyuran Chemexpress) and ethyl 2-bromobutyrate. The crude product was purified by flash chromatography (EtOAc/heptane gradient from 5:95 to 25: 75) to give (2S) -ethyl 2- { [ (1R) -1- (4-cyanophenyl) ethyl ] amino } butanoate (second eluting compound) as a colorless oil (345 mg, 19%) and ethyl (2R) -2- { [ (1R) -1- (4-cyanophenyl) ethyl ] amino } butanoate (first eluting compound) as a colorless oil (460 mg, 26%).

(2S)-2-{[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ] Amino group } Butyric acid ethyl ester : ¹H NMR (DMSO-d₆300 MHz) δ 7.76 (d, J = 8.4 Hz, 2H), 7.52 (d, J = 8.4 Hz, 2H), 3.86 (m, 2H), 3.05 (m,1H), 2.42 (m,1H), 1.51 (m, 2H), 1.23 (d, J = 6.7 Hz, 3H), 1.07 (t, J =7.1 Hz, 3H), 0.82 (t, J =7.4 Hz, 3H). LC/MS (method B) 261.0 (M + H) ⁺。

(2R)-2-{[(1R)-1-(4- Cyanophenyl group ) Ethyl radical ] Amino group } Butyric acid ethyl ester : ¹H NMR (DMSO-d₆300 MHz) δ 7.78 (d, J = 8.3 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 4.09 (q, J =7.01 Hz, 2H), 3.72 (m, 2H), 2.51 (m, 1H), 1.50 (m, 2H), 1.22 (d, J = 6.5 Hz, 3H), 1.17 (t, J =7.1 Hz, 3H), 0.82 (t, J =7.5 Hz, 3H). LC/MS (method B) 261.0 (M + H)⁺. HPLC (method A) Rt 2.17 min (purity: 99%).

Step (ii) of 2: (2S)-2-[((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Amino group ] Butyric acid ethyl ester

Following general procedure 1, starting from (2S) -2- { [ (1R) -1- (4-cyanophenyl) ethyl]Amino } butyric acid ethyl ester the title compound was prepared initially and isolated as a colorless oil (330 mg, 85%).¹H NMR (DMSO-d₆300 MHz) δ 9.57 (s, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H), 5.76 (brs, 2H), 3.95-3.85 (m, 2H), 3.71 (m, 1H), 3.07 (m, 1H), 2.19 (m, 1H), 1.55 (m, 2H), 1.24 (d, J = 6.6 Hz, 3H), 1.10 (t, J =7.1 Hz, 3H), 0.82 (t, J =7.4 Hz, 3H). LC/MS (method B) 294.0 (M + H)⁺。

Intermediates 13: (2R)-2-[((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Amino group ] Butyric acid ethyl ester

Following general procedure 1, starting from (2R) -2- { [ (1R) -1- (4-cyanophenyl) ethyl]Ethyl amino } butanoate (for its synthesis see intermediate 12, step 1) the title compound was prepared starting and isolated as a solid (480 mg, 93%). ¹H NMR (DMSO-d₆300 MHz) δ 9.57 (s, 1H), 7.59 (d, J = 8.2 Hz, 2H), 7.29 (d, J = 8.2 Hz, 2H), 5.76 (brs, 2H), 4.10 (q, 2H, J =7.1 Hz), 3.63 (m, 1H), 2.72 (m, 1H), 2.34 (m, 1H), 1.49 (m, 2H), 1.25-1.13 (m, 6H), 0.81 (t, J =7.5 Hz, 3H). LC/MS (method B) 294.0 (M + H)⁺。

Intermediates 14: 2-({3-[ Amino group ( Oximino radical ) Methyl radical ]-5- Fluorobenzyl radical } Amino group )-4- Methylpentanoic acid ethyl ester

Step (ii) of 1: 3-( Bromomethyl radical )-5- Fluorobenzonitriles

3-fluoro-5-methylbenzonitrile (Hognda tracing; 100 g, 0.74 mol) was dissolved in acetonitrile (1L) at 25 ℃ under a nitrogen atmosphere. N-bromosuccinimide (105 g, 0.59 mol) and AIBN (2.4 g, 0.014 mol) were added and the reaction mixture was heated at 70 ℃ for 1 hour 20 minutes. The reaction mixture was cooled to 25 ℃ and concentrated. The residue was diluted, cooled to 0-5 ℃ and stirred at the same temperature for 15 minutes. The precipitated succinimide was filtered, and the filtrate was concentrated to give the crude product as a yellow oil (90 g). It was dissolved in petroleum ether (200 mL) and cooled to-20 ℃ and stirred for 30 minutes. The precipitated solid was filtered and dried to give the title product as a white solid.¹H NMR (DMSO-d₆(ii) a 400 MHz). delta.7.83 (m, 2H), 7.73 (m, 1H), 4.72 (s, 2H). HPLC (method A) Rt 4.17 min (purity: 99.4%).

Step (ii) of 2: 2-[(3- Cyano radical -5- Fluorobenzyl radical ) Amino group ]-4- Methyl amyl acid tert-butyl ester

To 3- (bromomethyl) -5-fluorobenzonitrile (4.0 g, 0.02 mol) in dry DMF (40 mL) was added sodium bicarbonate (3.9 g, 0.046 mol) and ethyl-D-leucine tert-butyl ester hydrochloride (4.2 g, 0.019 mol). The reaction mixture was stirred at room temperature for 16 hours. Diluting with water, and adding ethyl acetateAnd (4) ester extraction. The organic phase was washed with water, brine, dried, and concentrated. The crude product was purified by chromatography using petroleum ether/ethyl acetate as eluent to give the title compound as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz): δ7.68 (d, J = 9.5 Hz, 1H), 7.63 (s, 1H), 7.53 (d, J = 9.8 Hz, 1H), 3.78 (d, 1H), 3.64 (d, 1H), 2.97 (d, 1H), 2.60 (brs, 1H), 1.97-1.71 (m, 1H),1.36 (m, 11H), 0.86 (d, 3H), 0.80 (d, 3H)。

Step (ii) of 3: 2-({3-[ Amino group ( Oximino radical ) Methyl radical ]-5- Fluorobenzyl radical } Amino group )-4- Methylpentanoic acid ethyl ester

To a solution of 2- [ (3-cyano-5-fluorobenzyl) amino group in ethanol (30 mL)]To tert-butyl-4-methylpentanate (3.0 g, 9.4 mmol) was added hydroxylamine (1.2 mL, 0.019 mol) and it was stirred at 0 ℃ for 16 hours. The reaction mixture was diluted with water (250 mL) and extracted with ethyl acetate. The organic layer was separated, washed with brine, dried, and concentrated. The crude product was purified by chromatography using petroleum ether/ethyl acetate as eluent to give the title compound as a white solid (2.3 g, 70%). ¹H NMR (DMSO-d₆400 MHz δ 9.73 (s, 1H), 7.46 (s, 1H), 7.30 (d, J = 9.6 Hz, 1H), 7.13 (d, J = 9.6 Hz, 1H), 5.84 (s, 2H), 3.75 (d, 1H), 3.55 (d, 1H), 2.98 (s, 1H), 2.38 (brs, 1H), 1.76-1.69 (m, 1H), 1.40 (s, 9H), 1.37 (dd, 2H), 0.96 (d, 3H), 0.87 (d, 3H). LC/MS (method B) 354.3 (M + H)⁺. HPLC (method A) Rt 2.96 min (98.3% purity).

Intermediates 15: N-{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }-N- Methyl valine tert-butyl ester

Step (ii) of 1: 2- Amino group -3- Tert-butyl methylbutyrate

To a stirred solution of DL-valine (25 g, 0.213 mol) in tert-butyl acetate (250 mL) was added HClO in portions at 0 deg.C₄(64.2 g, 0.320 mol). The reaction mixture was stirred at room temperature for 36 h. It was diluted with water and extracted in ethyl acetate (500 mL). The organic layer was washed with 10% sodium bicarbonate solution (2X150 mL) over Na₂SO₄Dried and concentrated to give the title compound as a brown liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.31 (brs, 2H), 3.62(d, J = 4.4 Hz, 1H), 1.98 (d, J = 1.7 Hz, 1H), 1.44(s, 9H), 0.95 (m, 6H)。

Step (ii) of 2: 2-[(4- Cyano radical -2- Fluorobenzyl radical ) Amino group ]-3- Tert-butyl methylbutyrate

To a stirred solution of tert-butyl 2-amino-3-methylbutyrate (8.5 g, 0.05 mol) in dry DMF (50 mL) under nitrogen was added 4-cyano-2-fluoro-benzyl bromide (FluoroChem Ltd, 9.4 g, 0.044 mol) and NaHCO₃(10.2 g, 0.12 mol). The resulting mixture was stirred at room temperature for 16 h. Water (70 mL) was added and the desired product was extracted with ethyl acetate (2 × 100 mL). The organic layer was washed with water (3X100 mL) and Na ₂SO₄The solvent was dried and concentrated under vacuum. The resulting residue was purified by column chromatography using petroleum ether/ethyl acetate as eluent to give the title compound as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.77 (d, J = 10.2 Hz, 1H), 7.67 (t, J = 1.5 Hz, 1H), 3.85-3.79 (m, 2H), 2.72 (m, 1H), 2.49 (m, 1H), 1.81 (m, 1H), 1.37(s, 9H), 0.86 (m, 6H)。

Step (ii) of 3: 2-[(4- Cyano radical -2- Fluorobenzyl radical )( Methyl radical ) Amino group ]-3- Tert-butyl methylbutyrate

2- [ (4-cyano-2-fluorobenzyl) amino in DMF (20 mL) at 0 deg.C]Tert-butyl-3-methylbutyrate (7.2 g, 0.023 mol) was added dropwise to a stirred suspension of sodium hydride (2.2 g, 0.047 mol) in dry DMF (10 mL). The resulting mixture was stirred at room temperature for 20 min. Methyl iodide (6 mL, 0.094 mol) was then added dropwise at 0 ℃ and the mixture was stirred at room temperature for 3 hours. The reaction mixture was quenched in ice-water and extracted with ethyl acetate (250 mL). The organic layer was washed with water (3X100 mL) and Na₂SO₄Dried and concentrated under reduced pressure. The crude product was purified by column chromatography using petroleum ether and ethyl acetate as eluents to give the title compound as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.80 (d, J = 10 Hz, 1H), 7.69 (d, J = 7.9Hz, 2H), 7.60 (t, J = 7.4 Hz, 1H), 3.79 (d, J = 15 Hz, 1H), 3.64 (d, J = 15 Hz, 1H), 2.65 (d, J = 10.7 Hz, 1H), 2.18 (s, 3H), 1.44 (s, 9H), 0.92-0.83 (m, 6H)。

Step (ii) of 4: N-{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }-N- Methyl valine tert-butyl ester

To a stirred 2- [ (4-cyano-2-fluorobenzyl) (methyl) amino group]To a solution of tert-butyl (3.4 g, 0.011mol) 3-methylbutyrate in ethanol (30 mL) was added 50% aqueous hydroxylamine solution (1.4 mL, 0.021 mol). The mixture was stirred at-20 ℃ for 12 h. The reaction mixture was diluted with water and extracted in dichloromethane (100 mL) over Na ₂SO₄Dried and concentrated under reduced pressure. The solid was recrystallized from ethyl acetate (5 mL), filtered, and dried under vacuum,the title compound was obtained as a white solid.¹H NMR (DMSO-d₆400 MHz) δ 9.73(s, 1H), 7.49 (m, 1H), 7.38 (m, 2H), 5.85 (s, 2H), 3.71 (d, J =14.2 Hz, 1H), 3.57 (d, J =14.2 Hz, 1H), 2.66 (d, J =10.7 Hz, 1H), 2.18 (s, 3H),1.99-1.93 (m, 1H), 1.45 (s, 9H), 0.91-0.81 (m, 6H). LC/MS (method B) 354.3 (M + H)⁺. HPLC (method A) Rt 2.66 min (98.6% purity).

Intermediates 16: N-(1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methylglycine tert-butyl ester

Step (ii) of 1: 4-[1-( Methylamino radical ) Ethyl radical ] Benzonitrile

To THF (50 mL) was added 4-acetylbenzonitrile (5.0 g; 34 mmol), methylamine (20.5 mL; 2M; 41 mmol) and titanium (IV) isopropoxide (5.6 mL; 18.9 mmol). The mixture was stirred at room temperature overnight. Sodium borohydride (5.5 g; 145 mmol) was added and the reaction stirred at room temperature for 2 h. 25% ammonium hydroxide solution (100 mL) was added and the reaction was stirred at room temperature for 1 hour 30 minutes. Then filtered through a celite pad, washing the pad 3 times with EtOAc. The filtrate was poured into a separatory funnel and saturated NaHCO was used₃The organic phase was washed with brine, MgSO₄Drying, filtration and concentration under reduced pressure gave the title compound as a pale yellow oil (5.4 g; 97%). It was used in the next step without further purification.

Step (ii) of 2: N-[1-(4- Cyanophenyl group ) Ethyl radical ]-N- Methylglycine tert-butyl ester

According to general procedure 11, starting from 4- [1- (methylamino) ethyl]The title compound was prepared starting from benzonitrile and tert-butyl bromoacetate. It was isolated as a colorless oil.¹H NMR (DMSO-d₆300 MHz) δ 7.79 (d, J = 8.4 Hz, 2H), 7.54 (d, J = 8.2 Hz, 2H), 3.89 (q, J = 6.7 Hz, 1H), 3.27 (d, J = 16.7 Hz, 1H), 3.07 (d, J = 16.7 Hz, 1H), 2.21 (s, 3H), 1.41 (s, 9H), 1.26 (d, J = 6.8 Hz, 3H). HPLC (method A) Rt 2.83 min (purity: 96.8%).

Step (ii) of 3: N-(1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methylglycine tert-butyl ester

From N- [1- (4-cyanophenyl) ethyl according to general procedure 1]The title compound was prepared starting from tert-butyl-N-methylglycinate and isolated as a white powder (2.68 g, quant.).¹H NMR (DMSO-d₆300 MHz) 9.58 (s, 1H), 7.61 (d, J = 8.2 Hz, 2H), 7.30 (d, J = 8.3 Hz, 2H), 5.78 (s, 2H), 3.80 (q, J = 6.7 Hz, 1H), 3.23 (d, J = 16.8 Hz, 1H), 3.03 (d, J = 16.8 Hz, 1H), 2.22 (s, 3H), 1.41 (s, 9H), 1.26 (d, J = 6.8 Hz, 3H). LC/MS (method B) 308.2 (M + H)⁺. HPLC (method A) Rt 1.33 min (purity: 88.5%).

Intermediates 17: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group )-2- (iii) methyl ] propionic acid tert-butyl ester

Step (ii) of 1: 2-( Benzylamino group )-2- (iii) methyl ] propionic acid tert-butyl ester

The title compound was prepared following general procedure 10 starting from tert-butyl 2-bromoisobutyrate and benzylamine. It was separated as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.30-7.26 (m, 4H), 7.23-7.20 (m, 1H), 3.55 (s, 2H), 2.10 (s, 1H), 1.42 (s, 9H), 1.20 (s, 6H)。

Step (ii) of 2: 2- Amino group -2- (iii) methyl ] propionic acid tert-butyl ester

To a solution of tert-butyl 2- (benzylamino) -2-methylpropionate (20 g, 0.08 mol) in ethyl acetate (1L) was added the catalyst palladium on carbon (10%, 2.2 g). The resulting mixture was placed at 5 Kg/cm²The reaction was maintained at room temperature for 18h under hydrogen pressure. The reaction mixture was filtered. The filtrate was concentrated to give the title compound as a pale green liquid.¹H NMR (DMSO-d₆, 400 MHz) δ1.38 (brs, 2H), 1.14 (s, 9H), 1.14-1.11 (d, J=10.7Hz, 6H)。

Step (ii) of 3: 2-[(4- Cyano radical -2- Fluorobenzyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

The title compound was prepared following general procedure 10 starting from tert-butyl 2-amino-2-methylpropionate and 4-cyano-2-fluorobenzyl bromide. The title compound was isolated as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.78-7.75 (d, J = 10.0 Hz, 1H), 7.71-7.65 (m, 2H), 3.70-3.68 (d, J = 6.9 Hz, 2H), 2.55 (s, 1H), 1.44 (s, 9H), 1.19 (s, 6H)。

Step (ii) of 4: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group )-2- (iii) methyl ] propionic acid tert-butyl ester

From 2- [ (4-cyano-2-fluorobenzyl) amino group according to general procedure 1]The title compound was prepared starting from tert-butyl-2-methylpropionate and isolated as a white solid.¹H NMR (DMSO-d₆400 MHz) delta 9.71 (s, 1H), 7.49-7.38 (m, 3H), 5.85 (s, 2H), 3.61 (s, 2H), 2.29 (s, 1H), 1.41(s, 9H), 1.20 (s, 6H). LC/MS (method B) 326.3 (M + H) ⁺. HPLC (method a) Rt: 2.25 min (purity: 99.4%).

Intermediates 18: (1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Carbamic acid tert-butyl ester

Step (ii) of 1: [1-(4- Cyanophenyl group ) Ethyl radical ] Carbamic acid tert-butyl ester

A solution of 4- (1-amino-ethyl) -benzonitrile (Ukrorgsynthsis Ltd, 2 g; 11 mmol) and N, N-diisopropylethylamine (2.2 mL; 13.1 mmol) in ACN (30 mL) was prepared. A solution of di-tert-butyl dicarbonate (2.9 g; 13.1 mmol) in ACN (10 mL) is added at 0 ℃ and the resulting solution is stirred at room temperature overnight. The solvent was concentrated, the reaction mixture was diluted with MTBE (100 mL), and H was added₂O (2 × 50 mL) and brine (50 mL). The aqueous layer was extracted with MTBE (100 mL). The organic layers were combined and washed with Na₂SO₄Dried and evaporated. Purification by flash chromatography gave the title compound as a yellow paste(2.2 g, 82%). LC/MS (method B) 246.9 (M + H)⁺, 245.2 (M-H)^-. HPLC (method A) Rt 1.20 min (purity: 100%).

Step (ii) of 2: (1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Carbamic acid tert-butyl ester

According to general procedure 1, from [1- (4-cyanophenyl) ethyl]The title compound was prepared starting from tert-butyl carbamate and isolated as a white powder (2.3 g, 94%). LC/MS (method B) 280.3 (M + H)⁺. HPLC (method A) Rt 2.11 min (purity: 99.6%).

Intermediates 19: N-(1-{3-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl alanine methyl ester

Step (ii) of 1: 3-[1-( Methylamino radical ) Ethyl radical ] Benzonitrile

The title compound was prepared following intermediate 16, the procedure used in step 1, starting from 3-acetylbenzonitrile. It was isolated as a colorless oil (1 g; 92%). HPLC (method A) Rt 1.54 min (purity: 74%).

Step (ii) of 2: N-[1-(3- Cyanophenyl group ) Ethyl radical ]-N- Methyl alanine methyl ester

This compound was prepared according to general procedure 11 starting from 3- [1- (methylamino) ethyl ] benzonitrile and methyl 2-bromopropionate to give the title compound as a colorless oil. HPLC (method A) Rt 1.54 min (purity: 74%).

Step (ii) of 3: N-(1-{3-[( Hydroxyamino group )( Imino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl alanine methyl ester

From N- [1- (3-cyanophenyl) ethyl according to general procedure 1]The title compound was prepared starting from methyl-N-methylalaninate and isolated as a colorless oil (330 mg, 88%). LC/MS (method B) 280.0 (M + H)⁺。

Intermediates 20: N-(1-{3-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl Glycine (methyl tert-butyl) ester

Step (ii) of 1: N-[1-(3- Cyanophenyl group ) Ethyl radical ]-N- Methyl alanine methyl ester

The title compound was prepared following general procedure 11 starting from 3- [1- (methylamino) ethyl ] benzonitrile (intermediate 19, step 1) and tert-butyl bromoacetate. It was isolated as a colorless oil. HPLC (method A) Rt 2.42 min (purity: 100%).

Step (ii) of 2: N-(1-{3-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl Glycine (methyl tert-butyl) ester

From N- [1- (3-cyanophenyl) ethyl according to general procedure 1]The title compound was prepared starting from methyl-N-methylalaninate and isolated as a colorless viscous oil (1.43 g, 98%). LC/MS (method B) 308.1 (M + H)⁺. HPLC (method A): 1.78 min (purity: 78.4%).

Intermediates 21: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

Step (ii) of 1: 3- Fluorine -4-[( Methylamino radical ) Methyl radical ] Benzonitrile

A solution of 4-cyano-2-fluoro-benzyl bromide (fluoroChem ltd, 16 g, 74 mmol) in dry THF (50 mL) was added to 40% aqueous methylamine solution (700 mL) at 5-10 deg.C and stirred for 5 min. The reaction mixture was extracted with DCM and Na₂SO₄The solvent was dried and concentrated in vacuo to give (12.0 g, 95%) the title compound as a brown liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.79-7.75 (m, 1H), 7.68-7.61 (m, 2H), 3.71 (s, 2H), 2.24 (s, 3H)。

Step (ii) of 2: 2-[(2- Fluorine -4- Isocyanobenzyl radical )( Methyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

From 3-fluoro-4- [ (methylamino) methyl according to general procedure 11]The title compound was prepared starting from benzonitrile and tert-butyl 2-bromoisobutyrate. It was isolated as a colorless oil.¹H NMR (DMSO-d₆, 400 MHz) δ7.78-7.76 (d, J = 10.3Hz, 1H), 7.73-7.67 (m, 2H), 3.68 (s, 2H), 2.13 (s, 3H), 1.41 (s, 9H), 1.27 (s, 6H)。

Step (ii) of 3: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

From 2- [ (2-fluoro-4-isocyanobenzyl) (methyl) amino according to general procedure 1 ]The title compound was prepared starting from tert-butyl-2-methylpropionate and isolated as a white solid.¹H NMR (DMSO-d₆400 MHz) delta 9.71 (s, 1H), 7.49-7.37 (m, 3H), 5.85 (s, 2H), 3.59 (s, 2H), 2.12 (s, 3H), 1.42 (s, 9H), 1.28 (s, 6H). LC/MS (method B) 340.3 (M + H)⁺. HPLC (method a) Rt: 2.14 min (purity: 98.1%).

Intermediates 22: N-(1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

Step (ii) of 1: N-[1-(4- Cyanophenyl group ) Ethyl radical ] Glycine tert-butyl ester

4-Acetylbenzonitrile (500 mg; 3.4 mmol; 1 eq.) and tert-butyl glycinate (588 mg; 4.5 mmol) were dissolved in toluene (10 mL). AcOH (103. mu.l; 1.7 mmol) was added. The mixture was stirred overnight with a Dean-Stark separator under reflux. Evaporating the solvent and reacting the resulting N- [1- (4-cyanophenyl) ethylene]Glycine tert-butyl ester (889 mg; 3.4 mmol) was dissolved in MeOH (18 mL). Sodium borohydride (390 mg; 10.3 mmol) was added in portions, giving an exothermic reaction (reflux). The reaction was stirred at room temperature overnight. The solvent was evaporated. The crude residue was suspended in EtOAc and washed with NH₄Saturated solution of Cl, NaHCO₃Saturated solution, brine, and over MgSO₄And (5) drying. After evaporation of the solvent, the resulting yellow oil was purified by flash chromatography to give the title compound as colorless oil. LC/MS (method B) 261.0 (M + H) ⁺. HPLC (method A): 2.77 min (purity: 98.2%).

Step (ii) of 2: N-(1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

From N- [1- (4-cyanophenyl) ethyl according to general procedure 1]The title compound was prepared starting with tert-butyl glycinate and isolated as a colorless foam (290 mg, 94%). LC/MS (method B): 294.1 (M + H)⁺. HPLC (method A): 1.31 min (purity: 100%)

Intermediates 23: N-((1S)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl glycine tert-butyl ester and intermediates 24: N-((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methylglycine tert-butyl ester

Step (ii) of 1: N-[1-(4- Cyanophenyl group ) Ethyl radical ]-N- Methylglycine tert-butyl ester

To THF (50 mL) was added 4-acetylbenzonitrile (5.0 g; 34 mmol), methylamine (21 mL; 2.00M solution in THF; 42 mmol) and titanium isopropoxide (5.6 mL; 19 mmol). The mixture was stirred at room temperature overnight. Sodium borohydride (5.5 g; 145 mmol) was added and the reaction mixture was stirred at room temperature for 2 h. 25% ammonium hydroxide solution (100 mL) was added and the reaction mixture was stirred at room temperature for 1 hour 30 minutes. Then filtered through a celite pad, washing the pad with EtOAc. With saturated NaHCO₃The organic phase was washed with brine, MgSO₄Dried, filtered and concentrated under reduced pressure. The crude product is then dissolved in CH ₃CN (41 mL). Potassium carbonate (10.3 g; 74 mmol) and tert-butyl bromoacetate (5.5 mL; 37 mmol) were added at room temperature, and the reaction mixture was stirred at room temperature for 1 hour 30 minutes. EtOAc is added and the organic phase is washed with water and brine over MgSO₄Drying, filtration, and concentration gave an oil which was purified by flash chromatography to give the title compound as a colorless oil.¹H NMR (DMSO-d₆300 MHz) δ 7.79 (d, J = 8.4 Hz, 2H), 7.54 (d, J = 8.2 Hz, 2H), 3.89 (q, J = 6.7 Hz, 1H), 3.27 (d, J = 16.7 Hz, 1H), 3.07 (d, J = 16.7 Hz, 1H), 2.21 (s, 3H), 1.41 (s, 9H), 1.26 (d, J = 6.8 Hz, 3H). LC/MS (method B) 275.1 (M+H)⁺. HPLC (method A) Rt 2.83 min (purity: 96.8%).

Step (ii) of 2: N-((1S)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methylglycine tert-butyl ester and N-((1R)-1-{4-[ amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methylglycine tert-butyl ester

Following general procedure 1, starting from N- [1- (4-cyanophenyl) ethyl ] -N-methylglycine tert-butyl ester obtained in step 1 (1.4 g; 5.1 mmol), two title compounds were prepared giving 1.6 g (quantitative) as white solids. This was purified by chiral HPLC on a Chiralcell OJ-H250 mmX20mmX5uM column eluting with isopropanol/heptane (v: v = 15:85) at a flow rate of 8.0 mL/min to give N- ((1S) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylglycine tert-butyl ester (second eluting compound) as a white solid (677 mg; 48%) and N- ((1R) -1- {4- [ amino (hydroxyimino) methyl ] phenyl } ethyl) -N-methylglycine tert-butyl ester (first eluting compound) as a white solid (606 mg; 43%).

N- ((1S) -1- {4- [ amino (hydroxyimino) methyl]Phenyl } ethyl) -N-methylglycine tert-butyl ester:¹H NMR (DMSO-d₆300 MHz) δ 9.58 (s, 1H), 7.61 (d, J = 8.3 Hz, 2H), 7.30 (d, J = 8.3 Hz, 2H), 5.78 (s, 2H), 3.80 (q, J = 6.7 Hz, 1H), 3.23 (d, J = 16.7 Hz, 1H), 3.03 (d, J = 16.7 Hz, 1H), 2.22 (s, 3H), 1.41 (s, 9H), 1.26 (d, J = 6.7 Hz, 3H). LC/MS (method B) 308.2 (M + H)⁺. HPLC (method B) Rt 12.55 min (purity: 100.0%).-52.8 (c 0.90, EtOH)。

N- ((1R) -1- {4- [ amino (hydroxyimino) methyl]Phenyl } ethyl) -N-methylglycine tert-butyl ester:¹H NMR (DMSO-d₆300 MHz) δ 9.58 (s, 1H), 7.61 (d, J = 8.3 Hz, 2H), 7.30 (d, J = 8.3 Hz, 2H), 5.78 (s, 2H), 3.80 (q, J = 6.7 Hz, 1H), 3.23 (d, J = 16.7 Hz, 1H), 3.03 (d, J = 16.7 Hz, 1H), 2.22 (s, 3H), 1.41 (s, 9H), 1.26 (d, J = 6.7 Hz, 3H). LC/MS (method A) 308.2 (M + H)⁺. HPLC (method B) Rt 8.99 min (purity: 100.0%).50.5 (c 1.13, EtOH)。

Intermediates 25: N-(1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl alanine methyl ester

Step (ii) of 1: N-[1-(4- Cyanophenyl group ) Ethyl radical ]-N- Methyl alanine methyl ester

To THF (50 mL) was added 4-acetylbenzonitrile (5.0 g; 34 mmol), methylamine (21 mL; 2.00M solution in THF; 42 mmol) and titanium isopropoxide (5.6 mL; 19 mmol). The mixture was stirred at room temperature overnight. Sodium borohydride (5.5 g; 145 mmol) was added and the reaction mixture was stirred at room temperature for 2 h. 25% ammonium hydroxide solution (100 mL) was added and the reaction mixture was stirred at room temperature for 1 hour 30 minutes. Then filtered through a celite pad, washing the pad with EtOAc. With saturated NaHCO ₃The organic phase was washed with brine, MgSO₄Dried, filtered and concentrated under reduced pressure. Then will beThe crude product is dissolved in CH₃CN (15 mL). Potassium carbonate (3.8 g; 27 mmol) and methyl 2-bromopropionate (1.5 mL; 13.7 mmol) were added at room temperature, and the reaction mixture was stirred at room temperature for 1 hour 30 minutes. EtOAc is added and the organic phase is washed with water and brine over MgSO₄Dried, filtered, and concentrated. The resulting oil was purified by flash chromatography to give the title compound as a colorless oil.¹H NMR (DMSO-d₆300 MHz) δ 7.83-7.76 (m, 2H), 7.54 (t, J = 8.7 Hz, 2H), 3.94-3.79 (m, 1H), 3.70-3.63 (m, 0.5H), 3.63-3.59 (m, 3H), 3.35-3.26 (m, 0.5H), 2.20 (s, 1.5H), 2.09 (s, 1.5H), 1.30-1.23 (m, 3H), 1.20 (d, J = 7.0 Hz, 1.5H), 1.12 (d, J = 7.0 Hz, 1.5H). LC/MS (method B) 247.1 (M + H)⁺. HPLC (method A) Rt 1.51 min (purity: 70.1%).

Step (ii) of 2: N-(1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical )-N- Methyl alanine methyl ester

From N- [1- (4-cyanophenyl) ethyl according to general procedure 1]The title compound was prepared starting from methyl-N-methylalaninate. It was isolated as a colorless oil (113 mg; quantitative). LC/MS (method B) 280.1 (M + H)⁺。

Intermediates 26: N-(1-{3-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

Step (ii) of 1: N-[1-(3- Cyano radicalPhenyl radical ) Ethyl radical ] Glycine tert-butyl ester

4-Acetylbenzonitrile (1.0 g; 6.9 mmol) and tert-butyl glycinate (1.2 g; 9.0 mmol) were dissolved in toluene (20 mL). The mixture was stirred under reflux using a Dean-Stark separator. AcOH (207. mu.L; 3.4 mmol) was added and the mixture was further heated overnight. The solvent was evaporated. The crude product was dissolved in MeOH (35 mL). The solution was cooled to 0 ℃ and sodium borohydride (781 mg; 21 mmol) was added in portions. The reaction was stirred at 0 ℃ for 1h and at rt overnight. The solvent was evaporated. The crude residue was suspended in EtOAc and washed with NH₄Saturated solution of Cl, NaHCO₃The saturated solution was washed with brine and MgSO₄And (5) drying. After evaporation of the solvent, the resulting yellow oil was purified by flash chromatography to give the title compound 716 mg as colorless oil. LC/MS (method B) 261.0 (M + H)⁺. HPLC (method A) Rt 2.79 min (purity: 99.2%).

Step (ii) of 2: N-(1-{3-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Glycine tert-butyl ester

From N- [1- (3-cyanophenyl) ethyl obtained in step 1 according to general procedure 1]Glycine tert-butyl ester (716 mg; 2.8 mmol) was used to prepare the title compound. It was isolated as a colorless foam (646 mg; 80%). ¹H NMR (DMSO-d₆300 MHz) δ 9.58 (br s, 1H), 7.64-7.58 (m, 1H), 7.56-7.48 (m, 1H), 7.35-7.26 (m, 2H), 5.78 (s, 2H), 3.73 (q, J = 6.3 Hz, 1H), 3.06 (d, J = 17.1 Hz, 1H), 2.96 (d, J = 17.1 Hz, 1H), 1.38 (s, 9H), 1.25 (d, J = 6.3 Hz, 3H). LC/MS (method B): 294.1 (M + H)⁺. HPLC (method A) Rt 1.77 min (purity: 97.3%).

Intermediates 27: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group ) Tert-butyl butyrate

Step (ii) of 1: 2- (iv) Aminobutanoic acid tert-butyl ester

To a solution of tert-butyl 2-bromobutyrate (10 g, 44.8 mmol) in dry THF (50 mL) at room temperature was added 25% ammonium hydroxide solution (800 mL). The reaction was stirred at rt for 24 h. The reaction mixture was concentrated under vacuum at low temperature (35-38 ℃). Then extracted with DCM (3 × 200 mL). The combined organic phases are passed over Na₂SO₄Dried and concentrated in vacuo to give the title compound (6.2 g, 85%) as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ3.09-3.06 (t, J = 6.1 Hz, 1H), 1.75-1.71 (m, 2H), 1.63-1.57 (m, 2H) 1.46 (s, 9H), 0.96-0.92 (t, J= 7.5 Hz, 3H)。

Step (ii) of 2: 2-[(4- Cyano radical -2- Fluorobenzyl radical ) Amino group ] Tert-butyl butyrate

The title compound was prepared following general procedure 10, starting from 4-cyano-2-fluoro-benzyl bromide (Fluorochem, 8.2 g, 38 mmol) and tert-butyl 2-aminobutyrate (6.2 g, 38 mmol) (obtained in step 1). It was separated as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.79-7.76 (d, J = 10.2 Hz, 1H), 7.68-7.67 (t, J = 2.1 Hz, 2H), 3.84-3.79 (m, 1H) 3.71-3.66 (m, 1H), 2.95-2.91 (m, 1H), 2.52-2.48 (m, 1H), 1.58-1.53 (m, 2H), 1.49 (s, 9H), 0.87-0.83 (m, 3H)。

Step (ii) of 3: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group ) Tert-butyl butyrate

Following general procedure 1, from 2- [ (4-cyano-2-fluorobenzyl) amino group obtained in step 2]The title compound was prepared starting with tert-butyl butyrate and isolated as a white solid.¹H NMR (DMSO-d₆300 MHz) δ 9.72 (s, 1H), 7.49-7.38 (m, 3H) 5.85 (s, 2H), 3.75-3.72 (d, J = 14.0 Hz, 1H), 3.64-3.60 (d, J = 14 Hz, 1H), 2.94 (m, 1H), 2.31 (m, 1H), 1.55-1.50 (m, 2H), 1.39 (s, 9H), 0.86-0.82 (t, J = 7.4 Hz, 3H). LC/MS (method B) 326.3 (M + H)⁺. HPLC (method A) Rt 2.140 min (purity: 98.6%).

Intermediates 29: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

Step (ii) of 1: 2-[(2- Fluorine -4- Isocyanobenzyl radical )( Methyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

From 3-fluoro-4- [ (methylamino) methyl according to general procedure 11]The title compound was prepared starting from benzonitrile and tert-butyl 2-bromoisobutyrate. It was separated as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.77 (d, J = 10.3 Hz, 1H), 7.73-7.67 (m, 2H), 3.68 (s, 2H), 2.13 (s, 3H), 1.41 (s, 9H), 1.27 (s, 6H)。

Step (ii) of 2: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

From 2- [ (2-fluoro-4-isocyanobenzyl) (methyl) amino]The title compound was prepared starting from tert-butyl-2-methylpropionate and isolated as a white solid.¹H NMR (DMSO-d₆400 MHz) delta 9.71 (s, 1H), 7.49-7.37 (m, 3H), 5.85 (s, 2H), 3.59 (s, 2H), 2.12 (s, 3H), 1.42 (s, 9H), 1.28 (s, 6H). LC/MS (method A) 340.3 (M + H) +. HPLC (method A) Rt: 2.14 min (purity: 98%).

Intermediates 30: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ] (iii) Tert-butyl acrylate

Step (ii) of 1: 2-[(4- Cyano radical -2- Fluorobenzyl radical )( Methyl radical ) Amino group ] (iii) Tert-butyl acrylate

From 3-fluoro-4- [ (methylamino) methyl according to general procedure 11]The title compound was prepared starting from benzonitrile and tert-butyl 2-bromopropionate. It was separated as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.80-7.77 (t, J = 9.9 Hz, 1H), 7.68-7.61 (m, 2H), 3.83-3.70 (dd, 2H), 3.35-3.32 (t, J = 7.1 Hz, 1H), 2.19 (s, 3H), 1.42 (s, 9H), 1.19 (d, J = 7.1 Hz, 3H)。

Step (ii) of 2: 2-[{4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }( Methyl radical ) Amino group ] (iii) Tert-butyl acrylate

From 2- [ (4-cyano-2-fluorobenzyl) (methyl) amino group according to general procedure 2]The title compound was prepared starting from tert-butyl propionate and isolated as a white solid.¹H NMR (DMSO-d₆400 MHz) δ 9.73 (s, 1H), 7.49 (m, 1H), 7.40 (m, 2H), 5.86 (s, 2H), 3.75-3.62 (m, 2H), 3.33-3.10 (t, J = 8.7 Hz, 1H), 2.19 (s, 3H), 1.43 (s, 9H), 1.18 (d, J = 7.1 Hz, 3H). LC/MS (method A) 326.3 (M + H) +. HPLC (method a) Rt: 2.05 min (purity: 98.3%).

Intermediates 31: N-{4-[( Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }-O-( Tert-butyl radical )-N- Methylserine tert-butyl ester

Step (ii) of 1: 3- Tert-butoxy radical -2-[(4- Cyano radical -2- Fluorobenzyl radical ) Amino group ] (iii) Tert-butyl acrylate

To a stirred solution of 4-cyano-2-fluoro-benzyl bromide (5 g, 0.024 mol) and sodium bicarbonate (3.9 g, 0.047 mol) in dry DMF (50 mL) was added tert-butyl-o-L-serine tert-butyl ester hydrochloride (6 g, 0.024 mol). The mixture was stirred at room temperature for 12 h. The reaction was diluted with water (100 mL) and extracted in ethyl acetate (100 mL). The organic layer was washed with water (2X100 mL) and Na ₂SO₄Dried and concentrated under reduced pressure. The crude product was purified by column chromatography using petroleum ether and ethyl acetate as eluents to give the title compound as a colorless liquid (6.7 g, 82%).¹H NMR (DMSO-d₆, 400 MHz) δ7.77 (d, J=10.1 Hz, 1H), 7.67 (t, J=2.2 Hz, 2H), 3.90-3.74 (m, 2H), 3.47 (m, 1H), 3.41 (m, 1H), 3.16 (m, 1H), 1.43 (s, 9H), 1.17 (s, 9H)。

Step (ii) of 2: 3- Tert-butoxy radical -2-[(4- Cyano radical -2- Fluorobenzyl radical )( Methyl radical ) Amino group ] (iii) Tert-butyl acrylate

To a stirred suspension of sodium hydride (1.8 g, 0.038 mol) in dry DMF (10 mL) at 0 deg.C was added 3-tert-butoxy-2- [ (4-cyano-2-fluorobenzyl) amino group in DMF (20 mL) dropwise]Tert-butyl propionate (6.7 g, 0.019 mol). The resulting mixture was stirred at room temperature for 20 min. Methyl iodide (3.5 mL, 0.0057 mol) was then added dropwise at 0 ℃ and the mixture was stirred at room temperature for 3 hours. The reaction mixture was quenched in ice water and extracted with ethyl acetate (250 mL). The organic layer was washed with water (3 × 100 mL), dried over sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography using petroleum ether and ethyl acetate as eluents to give the title compound as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.78 (d, J = 9.8 Hz, 1H), 7.67 (t, J = 2.5 Hz, 2H), 3.92 (d, J = 15.2 Hz, 1H), 3.81-3.70 (d, J = 15.0 Hz, 1H), 3.63 (m, 1H), 3.52 (m, 2H), 2.26 (s, 3H), 1.42 (s, 9H), 1.08 (s, 9H)。

Step (ii) of 3: N-{4-[( Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical }-O-( Tert-butyl radical )-N- Methylserine tert-butyl ester

From 3-tert-butoxy-2- [ (4-cyano-2-fluorobenzyl) (methyl) amino group according to general procedure 2]The title compound was prepared starting from tert-butyl propionate and isolated as a white solid. ¹H NMR (DMSO-d₆400 MHz) δ 9.72 (s, 1H), 7.49 (d, J = 8 Hz, 1H), 7.40 (m, 2H), 5.85 (s, 2H), 3.80 (d, J = 14.1 Hz, 1H), 3.71-3.63 (m, 2H), 3.47 (m, 1H), 3.26 (t, J = 5.5 Hz, 1H), 2.25 (s, 3H), 1.43 (s, 9H), 1.09 (s, 9H). LC/MS (method A) 398.0 (M + H)⁺. HPLC (method A) Rt 3.13 min (purity 94.9%).

Intermediates 32: 2-({3-[ Amino group ( Oximino radical ) Methyl radical ]-5- Fluorobenzyl radical } Amino group )-2- (iii) methyl ] propionic acid tert-butyl ester

Step (ii) of 1: 3-( Aminomethyl group )-5- Fluorobenzonitriles

To 3- (bromomethyl) -5-fluorobenzonitrile (intermediate 14, step 1, 10 g, 0.47 mol) in THF (50 mL) was added 25% ammonium hydroxide solution (2.0L). The resulting mixture was stirred at 85 ℃ for 2 h. It was then cooled to room temperature and extracted with dichloromethane. The organic layer was washed with water, brine, dried, and concentrated to give the title compound as a pale yellow liquid (5 g, 71%).¹H NMR (DMSO-d₆400 MHz). delta.7.6 (d, J = 6.6 Hz, 1H), 7.57 (t, J = 0.6 Hz, 1H), 7.55 (s, 1H), 3.74 (d, 2H), 1.97 (s, 2H). LC/MS (method A) 151.0 (M + H)⁺. HPLC (method a) Rt: 0.88 min (purity 98%).

Step (ii) of 2: 2-[(3- Cyano radical -5- Fluorobenzyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

The title compound was prepared according to general procedure 11, starting from 3- (aminomethyl) -5-fluorobenzonitrile and 2-bromoisobutyrate to give the title compound as a pale yellow liquid (5 g, 91%). ¹H NMR (DMSO-d₆400 MHz) delta 7.67 (d, J = 7 Hz, 2H), 7.57 (d, J = 9.5 Hz, 1H), 3.67 (d, 2H), 2.67 (t, 1H), 1.40 (s, 9H), 1.19 (s, 6H). LC/MS (method A) 293.2 (M + H)⁺. HPLC (method a) Rt: 2.29 min (purity: 99.7%).

Step (ii) of 3: 2-({3-[ Amino group ( Oximino radical ) Methyl radical ]-5- Fluorobenzyl radical } Amino group )-2- (iii) methyl ] propionic acid tert-butyl ester

From 2- [ (3-cyano-5-fluorobenzyl) amino group according to the general procedure]The title compound was prepared starting from tert-butyl-2-methylpropionate and isolated as a white solid (5.1 g, 92%).¹H NMR (DMSO-d₆400 MHz δ 9.72 (s, 1H), 7.49 (s, 1H), 7.29 (d, J = 10.1 Hz, 1H), 7.15 (d, J = 9.7 Hz, 1H), 5.85 (s, 2H), 3.60 (d, 2H), 2.39 (d, 1H), 1.41 (s, 9H), 1.20 (s, 6H). LC/MS (method A) 326.3 (M-H)^-. HPLC (method A) Rt 2.26 min (purity: 98.7%).

Intermediates 33: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group )-3- Tert-butyl methylbutyrate

Step (ii) of 1: 2-[(4- Cyano radical -2- Fluorobenzyl radical ) Amino group ]-3- Methyl butyric acid -1- Tert-butyl ester

To a stirred solution of tert-butyl 2-amino-3-methylbutyrate (Bachem, 6.3 g, 0.036 mol) in dry DMF (50 mL) under nitrogen was added 4-cyano-2-fluoro-benzyl bromide (6.2 g, 0.029 mol) and NaHCO₃(6.09g, 0.073 mol). The resulting mixture was stirred at room temperature for 16 h. Water (70 mL) was added and extracted with ethyl acetate (2 × 100 mL). The organic layer was washed with water (3X100mL) over Na ₂SO₄The solvent was dried and concentrated under vacuum. The resulting residue was purified by column chromatography using petroleum ether/ethyl acetate as eluent to give the title compound as a colorless liquid.¹H NMR (DMSO-d₆, 400 MHz) δ7.78 (d, J = 10.2 Hz, 1H), 7.67 (t, J = 1.5 Hz, 2H) 3.81 (m, 2H), 2.71 (m, 1H), 2.49 (m 1H), 1.81 (m, 1H), 1.37 (s, 9H), 0.88-0.84 (m, 6H)。

Step (ii) of 2: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group )-3- Tert-butyl methylbutyrate

From 2- [ (4-cyano-2-fluorobenzyl) amino group according to the general procedure]The title compound was prepared starting from tert-butyl-3-methylbutyrate and isolated as a white solid. LC (liquid Crystal)/MS (method A) 340.3 (M + H)⁺. HPLC (method a) Rt: 2.47 min (purity: 98.7%).

Intermediates 34: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group )-2- (iii) methyl ] propionic acid tert-butyl ester

Step (ii) of 1: 2-[(4- Cyano radical -2- Fluorobenzyl radical ) Amino group ]-2- (iii) methyl ] propionic acid tert-butyl ester

To a stirred solution of tert-butyl 2-amino-2-methylpropionate (Bachem, 7.9 g, 0.05 mol) in dry DMF (50 mL) under nitrogen was added 4-cyano-2-fluorobenzyl bromide (10.6 g, 0.05 mol) and NaHCO as solids₃(8.3 g, 0.099 mol). The reaction mixture was stirred at 70 ℃ for 16 h. Water (70 mL) was added and extracted with ethyl acetate (100 mL). The organic layer was washed with water (3X100 mL) and Na₂SO₄The solvent was dried and concentrated under vacuum. The residue was purified by column chromatography using silica gel (60-120 mesh) and petroleum ether/ethyl acetate as eluent to give the title compound as a colorless liquid. ¹H NMR (DMSO-d₆, 400 MHz) δ7.78 (d, J = 10.0 Hz, 1H), 7.71-7.65 (m, 2H), 3.69 (d, J = 6.9 Hz, 2H), 2.55 (s, 1H), 1.44 (s, 9H), 1.19 (s, 6H)。

Step (ii) of 2: 2-({4-[ Amino group ( Oximino radical ) Methyl radical ]-2- Fluorobenzyl radical } Amino group )-2- (iii) methyl ] propionic acid tert-butyl ester

From 2- [ (4-cyano-2-fluorobenzyl) amino group according to the general procedure]The title compound was prepared starting from tert-butyl-2-methylpropionate and isolated as a white solid.¹H NMR (DMSO-d₆400 MHz) delta 9.71 (s, 1H), 7.49-7.38 (m, 3H), 5.85 (s, 2H), 3.61 (s, 2H), 2.29 (s, 1H), 1.41(s, 9H), 1.20 (s, 6H). LC/MS (method A) 326.3 (M + H)⁺. HPLC (method a) Rt: 2.26 min (purity: 99.4%).

Intermediates 35: 2-((1-(4-(N'- Hydroxycarbamimidoyl ) Phenyl radical )-2- Methoxy ethyl radical )( Methyl radical ) Amino group ) Acetic acid tert-butyl ester

Step (ii) of 1: 4-(2- Methoxyacetyl group ) Benzonitrile (a), (b), (c), (d 4-(2-methoxyacetoyl)benzonitrile ）

Isopropyl magnesium chloride (2M in THF; 3.0 mL; 6.0 mmol) was added to an ice-cooled solution of 4-iodobenzonitrile (1.2 g, 5.0 mmol) in anhydrous THF (10 mL). The solution was stirred at this temperature for 10 minutes and then cooled (-78 ℃ C.) was added dropwiseN-2-dimethoxy-N-a solution of methylacetamide (0.998 g, 7.50 mmol) in anhydrous THF (5 mL). The reaction mixture was stirred at this temperature for 1 hour, then at room temperature for 1 hour. The reaction mixture was then treated with 10% aqueous potassium hydrogen sulfate and extracted with DCM. The organic phase was passed through a hydrophobic frit filter plate and evaporated under reduced pressure. The residue was purified by flash chromatography, eluting with isohexane/EtOAc (3:2), The title compound was obtained.¹H NMR (CDCl₃, 400 MHz) δ 8.06-8.04 (m, 2H), 7.80-7.77 (m, 2H), 4.67 (s, 2H), 3.50 (s, 3H)。

Step (ii) of 2: 2-((1-(4- Cyanophenyl group )-2- Methoxy ethyl radical )( Methyl radical ) Amino group ) Acetic acid tert-butyl ester

A solution of 4- (2-methoxyacetyl) benzonitrile (0.338 g; 1.93 mmol) and glycine tert-butyl ester (0.382 mg; 2.90 mmol) in DCM/MeOH (1: 1; 10 mL) was stirred at ambient temperature for 1 h. Sodium cyanoborohydride (0.182 g; 2.90 mmol) and acetic acid (0.166 mL; 2.90 mmol) were added to the reaction mixture, and the mixture was stirred for 18 hours. An aqueous solution of paraformaldehyde (0.787 mL) and sodium cyanoborohydride (0.182 g; 2.90 mmol) was added and the reaction mixture was stirred for an additional 72 hours. The reaction mixture was diluted with DCM and water and then passed through a hydrophobic frit filter plate. The filtrate was evaporated in vacuo. The residue was purified by flash chromatography, eluting with isohexane/EtOAc (3:2), to give the title compound.¹H NMR (CDCl₃, 400 MHz) δ 7.62-7.61 (m, 2H), 7.51-7.49 (m, 2H), 4.09-4.06 (m, 1H), 3.62 (m, 2H), 3.42-3.37 (m, 1H), 3.29 (s, 3H), 3.21-3.17 (m, 1H), 2.39 (s, 3H), 1.47 (s, 9H)。

Step (ii) of 3: 2-((1-(4-(N'- Hydroxycarbamimidoyl ) Phenyl radical )-2- Methoxy ethyl radical )( Methyl radical ) Amino group ) Acetic acid tert-butyl ester

A solution of tert-butyl 2- ((1- (4-cyanophenyl) -2-methoxyethyl) (methyl) amino) acetate (0.136 g; 0.45 mmol) and 50% aqueous hydroxylamine solution (0.137 mL; 2.24 mmol) in ethanol (2 mL) was heated at 80 ℃ for 18 h. The solvent was evaporated in vacuo. In the presence of DCM and water The residue was partitioned. The organic phase was passed through a hydrophobic frit filter plate and evaporated in vacuo to give the title compound (0.141 g, 93%).¹H NMR (CDCl₃, 400 MHz) δ 7.78 (d,J = 8.4 Hz, 2H), 7.59-7.57 (m, 2H), 6.70 (br s, 1H), 4.84 (br s, 2 H), 4.05-4.02 (m, 1H), 3.71-3.67 (m, 1H), 3.63-3.60 (m, 1H), 3.40-3.35 (m, 1H), 3.34 (s, 3H), 3.21-3.16 (m, 1H), 2.40 (s, 3H), 1.49 (s, 9H)。

Intermediates 36: 2-(2-( Tert-butyldimethylsilyloxy )-1-(4-(N'- Hydroxycarbamimidoyl ) Phenyl radical ) Ethylamino group ) Acetic acid tert-butyl ester

Step (ii) of 1: 4-(2- Hydroxyacetyl radical ) Benzonitrile

Trifluoroacetic acid (3.5 mL; 47.1 mmol) was added to 4-acetylbenzonitrile (3.40 g; 23.4 mmol) and [ bis (trifluoroacetoxy) iodide]Benzene (20.1 g; 46.7 mmol) in acetonitrile/water (5: 1; 300 mL). The reaction mixture was heated under reflux for 18 hours. The solvent was removed in vacuo and the residue partitioned between DCM and 10% aqueous potassium carbonate. The organic phase was passed through a hydrophobic frit filter plate and the solvent was evaporated in vacuo. The residue was purified by flash chromatography, eluting with isohexane/EtOAc (10:1 to 4:1), to give the title compound.¹H NMR (CDCl₃, 400 MHz) δ 8.04 (d,J = 8.4 Hz, 2H), 7.84 (d,J = 8.4 Hz, 2H), 4.91 (d,J = 2 Hz, 2H), 3.37 (t,J = 2.0 Hz, 2H)。

Step (ii) of 2: 2-(1-(4- Cyanophenyl group )-2- Hydroxy ethylamino ) Acetic acid tert-butyl ester

A solution of 4- (2-hydroxyacetyl) benzonitrile (0.730 g; 4.53 mmol) and tert-butyl glycinate (0.896 mg; 6.80 mmol) in DCM/MeOH (1: 1; 12 mL) was stirred at ambient temperature for 1 h. Sodium cyanoborohydride (0.342 g; 6.80 mmol) and acetic acid (0.389 mL; 6.80 mmol) were added to the reaction mixture, and the mixture was stirred for 18 hours. The reaction mixture was diluted with DCM and water and then passed through a hydrophobic frit filter plate. The filtrate was evaporated in vacuo. The residue was purified by flash chromatography, eluting with isohexane/EtOAc (3:2), to give the title compound. ¹H NMR (CDCl₃, 400 MHz) δ 7.65 (d,J = 8.4 Hz, 2H), 7.49 (d,J = 8.4 Hz, 2H), 3.88-3.85 (m, 1H), 3.76-3.72 (m, 1H), 3.59-3.55 (m, 1H), 3.32 (d,J = 17.6 Hz, 1H), 3.15 (d,J = 17.6 Hz, 1H), 1.49 (s, 9H)。

Step (ii) of 3: 2-(2-( Tert-butyldimethylsilyloxy )-1-(4- Cyanophenyl group ) Ethylamino group ) Acetic acid tert-butyl ester

Tert-butyldimethylsilyl chloride (t-butyldimethylsilyl chloride) (0.579 g; 3.84 mmol) and imidazole (0.261 g; 3.83 mmol) were added to a solution of tert-butyl 2- (1- (4-cyanophenyl) -2-hydroxyethylamino) acetate (0.709 g; 2.56 mmol) in DMF (10 mL). The reaction mixture was stirred at ambient temperature for 18 hours. The mixture was diluted with EtOAc and washed sequentially with water and brine. The organic phase was passed through a hydrophobic frit filter plate and the solvent was evaporated in vacuo. The residue was purified by flash chromatography, eluting with isohexane/EtOAc (10:1), to give the title compound.¹H NMR (CDCl₃, 400 MHz) δ 7.63-7.61 (m, 2H), 7.48 (d,J = 8.4 Hz, 2H), 3.86-3.83 (m, 1H), 3.66-3.55 (m, 2H), 3.23 (d, J = 16.8 Hz, 1H), 3.06 (d,J = 16.8 Hz, 1H), 2.53 (br s, 1H), 1.47 (s, 9H), 0.90 (s, 9H), 0.10 (s, 3H), 0.09 (s, 3H)。

Step (ii) of 4: 2-(2-( Tert-butyldimethylsilyloxy )-1-(4-(N'- Hydroxycarbamimidoyl ) Phenyl radical ) Ethylamino group ) Acetic acid tert-butyl ester

A solution of tert-butyl 2- (2- (tert-butyldimethylsilyloxy) -1- (4-cyanophenyl) ethylamino) acetate (0.496 g; 1.27 mmol) and 50% aqueous hydroxylamine solution (0.389 mL; 6.49 mmol) in ethanol (5 mL) was heated at 70 ℃ for 18 h. The solvent was evaporated in vacuo. The residue was partitioned between DCM and water. The organic phase was passed through a hydrophobic frit filter plate and evaporated in vacuo to give the title compound (0.566 g, 100%). ¹H NMR (CDCl₃, 400 MHz) δ 7.59 (d,J = 8.4 Hz, 2H), 7.39 (d,J = 8.0 Hz, 2H), 7.00 (br s, 1H), 4.84 (br s, 2H), 3.84-3.81 (m, 1H), 3.68-3.57 (m, 2H), 3.25 (d,J = 17.2 Hz, 1H), 3.08 (d,J = 16.8 Hz, 1H), 2.60 (br s, 1H), 1.44 (s, 9H), 0.88 (s, 9H), 0.03 (s, 3H), 0.01 (s, 3H)。

Intermediates 37: N'- Hydroxy radical -4-( Hydroxymethyl group ) Benzamidine

To a solution of 4- (hydroxymethyl) benzonitrile (11.05 g; 83.08 mmol) in EtOH (100 mL) was added hydroxylamine (27.4 mL; 415 mmol) (50% in water) and the mixture was heated to 74 ℃ for 16 h. The mixture was poured into a crystallization dish and the solvent was evaporated. The residue was washed with copious amounts of EtOAc, dry MeOH and dry MeCN, filtered through a hydrophobic frit filter plate, and the solvent was removed in vacuo to give the title compound,as a white solid (13.1 g, 95%).¹H NMR (DMSO-d₆, 400 MHz) δ 9.58 (s, 1H), 7.70-7.62 (m, 2H), 7.34 (d, J = 8.1 Hz, 2H), 5.79 (s, 2H), 5.23 (t, J = 5.6 Hz, 1H), 4.54 (d,J = 5.6 Hz, 2H)。

Intermediates 38: N'- Hydroxy radical -3-( Hydroxymethyl group ) Benzamidine

The title compound was prepared following the procedure described for intermediate 37 but starting from 3- (hydroxymethyl) benzonitrile (8.4 g; 63.4 mmol) to give the title compound as a white solid (9.2 g, 86%).¹H NMR (DMSO-d₆, 400 MHz) δ 9.63 (s, 1H), 7.67 (s, 1H), 7.58-7.54 (m, 1H), 7.35 (d,J= 4.7 Hz, 2H), 5.82 (s, 2H), 5.27 (t,J= 5.7 Hz, 1H), 4.54 (d,J= 5.7 Hz, 2H)。

Intermediates 39: N'- Hydroxy radical -4-( Hydroxyethyl group ) Benzamidine

The title compound was prepared following the procedure described for intermediate 37 but starting from 4- (hydroxyethyl) benzonitrile (2.4 g; 16.5 mmol) to give the title compound as a white solid (3.0 g, 99%).¹H NMR (DMSO-d₆400 MHz) δ 9.58 (s, 1H), 7.60 (d, J = 7.9 Hz, 2H), 7.24 (d, J = 7.9 Hz, 2H), 5.75 (s, 2H), 4.75-4.69 (m, 1H), 3.64 (d, J = 6.5 Hz, 2H), 2.80-2.72 (m, 2H). LC/MS (method B) 181 (M + H) ⁺. HPLC (method I) Rt 8.15 min (purity: 98.7%).

Intermediates 40: N'- Hydroxy radical -3-( Hydroxyethyl group ) Benzamidine

The title compound was prepared following the procedure described for intermediate 37 but starting from 3- (hydroxyethyl) benzonitrile (9.1 g, 6.2 mmol) to give the title compound as a white solid (1.1 g; 94%).¹H NMR (DMSO-d₆400 MHz) δ 9.59 (s, 1H), 7.55 (s, 1H), 7.51 (d, J = 7.7 Hz, 1H), 7.33-7.21 (m, 2H), 5.77 (s, 2H), 4.66 (t, J = 5.2 Hz, 1H), 3.65 (q, J = 6.2 Hz, 2H), 2.77 (t, J = 7.1 Hz, 2H). LC/MS (method B) 181 (M + H)⁺. HPLC (method I) Rt 8.26 min (purity: 99.9%).

Intermediates 41: ((1R)-1-{4-[ Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Carbamic acid tert-butyl ester

Step (ii) of 1: [(1R)-1-(4- Cyanophenyl group ) Ethyl radical ] Carbamic acid tert-butyl ester

A solution of 4- ((R) -1-amino-ethyl) -benzonitrile (81.30 g; 556.13 mmol; 1.0 eq.) in DCM (500 ml) was prepared in a 5-neck round-bottom flask (2.5L) equipped with a mechanical stirrer, thermometer and addition funnel placed under a nitrogen atmosphere and cooled at 5 ℃. Duration of timeA solution of di-tert-butyl dicarbonate (121.38 g; 556.13 mmol; 1.0 eq.) in DCM (400 ml) was added dropwise over 30 minutes, maintaining the temperature between 5 ℃ and 10 ℃. After about 20 minutes, a precipitate appeared, but later re-dissolved. After 2 hours, the cooling bath was removed. After 5.5 hours, 0.1N aqueous HCl (300 ml), water (300 ml) and water (250 ml)/NaHCO were used ₃The reaction mixture was washed with a mixture of saturated aqueous solution (50 ml). Drying (MgSO)₄) The organic layer was concentrated in vacuo to give the title compound as a pale yellow oil (139.43 g, quantitative yield). LC/MS (method B) 246.9 (M + H)⁺, 244.9 (M-H)^-. HPLC (method A) Rt 3.89 min (purity: 98.8%).

Step (ii) of ((1R)-1-{4-[2: Amino group ( Oximino radical ) Methyl radical ] Phenyl radical } Ethyl radical ) Carbamic acid tert-butyl ester

According to general procedure 1, from [ (1R) -1- (4-cyanophenyl) ethyl]The title compound was prepared starting from tert-butyl carbamate and isolated as a white powder (139.2 g, 89%). LC/MS (method B) 279.9 (M + H)⁺. HPLC (method A) Rt 2.10 min (purity: 99.7%).

Intermediates 42: {3-[ Amino group ( Oximino radical ) Methyl radical ] Benzyl radical } Methyl carbamic acid tert-butyl ester

Step (ii) of 1: 3-[( Methylamino radical ) Methyl radical ] Benzonitrile

In a 3-neck round-bottom flask (250 ml) placed under a nitrogen atmosphere, a solution of 3- (bromomethyl) benzonitrile (5 g; 25.5 mmol; 1 eq) in dry THF (20 ml) was added dropwise over 1 hour to methylamine (63.76 ml; 2M; 127.52 mmol; 5 eq) (2M solution in THF). The reaction mixture was stirred at room temperature for 2 hours. Some bibenzylation (7%) was observed. The reaction mixture was filtered to remove salts. The filtrate was evaporated under reduced pressure. The residue was dissolved with DCM and evaporated again to give the title compound as a yellow oil (3.78 g, quantitative yield). It was used in the next step without any further purification. LC/MS (method B) 146.8 (M + H) ⁺。

Step (ii) of 2: (3- Cyanobenzyl radical ) Methyl carbamic acid tert-butyl ester

In a round-bottomed flask (100 ml) placed under a nitrogen atmosphere, a crude 3- [ (methylamino) methyl group was prepared]A solution of benzonitrile (3.78 g; 23.79 mmol; 1 eq) in DCM (37.80 ml) was cooled in an ice bath, and a solution of di-tert-butyl dicarbonate (5.19 g; 23.79 mmol; 1 eq) in DCM (18.90 ml) was added over a period of 2 minutes. The reaction mixture was stirred between 0 ℃ and room temperature for 30 min. The reaction mixture was concentrated under reduced pressure to give a yellow oil (6.87 g). By flash chromatography (from 90:10 to 70:30 gAn alkane/EtOAc gradient) to give the title compound as a colorless oil (5.13 g, 88%).¹H NMR (DMSO-d₆300 MHz) δ 7.76 (ddd, J = 6.9, 1.9, 1.8, 1H), 7.65 (s, 1H), 7.62-7.48 (m, 2H), 4.41 (s, 2H), 2.79 (s, 3H), 1.62-1.16 (m, 9H). HPLC (method A) Rt 4.28min (purity: 100.0%).

Step (ii) of 3: {3-[(Z)- Amino group ( Oximino radical ) Methyl radical ] Benzyl radical } Methyl carbamic acid tert-butyl ester

The title compound was prepared according to general procedure 1 starting from tert-butyl (3-cyanobenzyl) methylcarbamate and isolated as a white powder (4.95 g, 87%).¹H NMR (DMSO-d₆300 MHz) δ 9.63 (s, 1H), 7.62-7.48 (m, 2H), 7.42-7.29 (m, 1H), 7.21 (d, J = 7.6, 1H), 5.79 (s, 2H), 4.38 (s, 2H), 2.76 (s, 3H), 1.59-1.24 (m, 9H). LC/MS (method B) 280.3 (M + H) ⁺. HPLC (method A) Rt 2.73 min. (purity: 99.9%).

Intermediates 43: [{3-[ Amino group ( Oximino radical ) Methyl radical ] Benzyl radical }( Methyl radical ) Amino group ] Acetic acid tert-butyl ester

Step (ii) of 1: [(3- Cyanobenzyl radical )( Methyl radical ) Amino group ] Acetic acid tert-butyl ester

To a stirred solution of sarcosine tert-butyl ester hydrochloride (8.1 g, 44.9 mmol) and triethylamine (17 mL, 122.4 mmol) in ACN (100 mL) was added 3- (bromomethyl) benzonitrile (8.0 g, 40.8 mmol) portionwise over 10 min at 0 ℃. After stirring at room temperature for 3 hours, the reaction mixture was poured into water and extracted with DCM. Then, the organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to give the title compound as a pale green liquid (9.0 g, 85%).¹H NMR (DMSO-d₆, 400 MHz) δ 7.72 (2H, m), 7.64 (1H, m), 7.54 (1H, m), 3.66 (2H, s), 3.18 (2H, s), 2.22 (3H, s), 1.41 (9H, s)。

Step (ii) of 2: [{3-[ Amino group ( Oximino radical ) Methyl radical ] Benzyl radical }( Methyl radical ) Amino group ] Acetic acid tert-butyl ester

According to general procedure 1, from [ (3-cyanobenzyl) (methyl) amino group]The title compound was prepared starting from tert-butyl acetate. It was obtained as a white powder (8.5 g, 84%).¹H NMR (DMSO-d₆400 MHz) delta 9.57 (1H, s), 7.59 (1H, s), 7.53 (1H, m), 7.30 (2H, m), 5.75 (2H, s), 3.61 (2H, s), 3.15 (2H, s), 2.23 (3H, s), 1.41 (9H, s). LC/MS (method B) 294.0 (M + H)⁺. HPLC (method A) Rt 3.31 min (purity: 97.5%).

Intermediates 44: (1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethylamine (ethylamine)

Step (ii) of 1: [(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Carbamic acid tert-butyl ester

A solution of intermediate 1 (10.03 g; 35.8 mmol; 1.0 eq.) and N-methylmorpholine (4.13 ml; 37.59 mmol; 1.05 eq.) in dry THF (100 ml) was prepared in a 3-neck round-bottom flask (250 ml) placed under a nitrogen atmosphere and cooled with an ice bath at 5 ℃. Isobutyl chloroformate (4.66 ml; 35.8 mmol; 1.0 equiv.) is added dropwise over a period of 2-3 minutes, maintaining the temperature between 5 ℃ and 9 ℃. After 1 hour at 5 ℃, intermediate 41 was added over 2-3 minutes. After 2.5 h, the reaction mixture was diluted with MTBE (200 ml) and washed with water (2X100 ml), 0.1N aqueous NaOH (100 ml), water (100 ml) and brine (100 ml). Drying (MgSO)₄) The organic layer was concentrated under vacuum to about 35 g (small amount of crystals appeared). The mixture was diluted successively with MTBE (30 ml, flash crystallization), pentane (45 ml), MTBE (15 ml) and pentane (15 ml). The precipitate was filtered off and washed with MTBE/pentane 1:1 (1X, 75 ml)) and pentane (2X). Drying under vacuum (40 ℃, 45 minutes). Isolation of [ (1R) -1- (4- { (Z) -amino [ ({ [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) ]Carbonyl } oxy) imino]Methyl } phenyl) ethyl]Tert-butyl carbamate as a white powder (17.55 g, 91%). LC/MS (method B): 542.3 (M + H)⁺。540.4 (M-H)^-. HPLC (method A) Rt 5.58min(purity: 99.6%).

In a round-bottom flask (500 ml) placed under a nitrogen atmosphere, [ (1R) -1- (4- { (Z) -amino [ ({ [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl)]Carbonyl } oxy) imino]Methyl } phenyl) ethyl]Tert-butyl carbamate (17 g; 31.39 mmol; 1.0 eq.) was suspended in toluene (170 ml) and then heated at 100 ℃. After overnight, the reaction mixture was concentrated under reduced pressure to give the title compound as a pale yellow oil (19.06 g, quantitative yield). This compound was used in the next step without further purification. LC/MS (method B) 582.4 (M + AcO)^-. HPLC (method A) Rt 6.57min (purity: 99.1%).

Step (ii) of 2: (1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethylamine (ethylamine)

In a round-bottomed flask (500 ml), prepared was [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl)]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]A solution of tert-butyl carbamate (19 g; 30.85 mmol; 1 eq.) in AcOH (95 ml). Then, hydrogen chloride (9.09 ml; 92.54 mmol; 3 equivalents) (32% aqueous solution) was added. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum to remove most of the AcOH. The residue was dissolved with MTBE (300 ml) and washed with 3N aqueous NaOH (300 ml), water (150 ml) and brine (150 ml). The aqueous layer was extracted with MTBE (100 ml). The combined organic layers were dried (MgSO) ₄) And concentrated to give the title compound as a pale yellow oil (13.13 g, quantitative yield).¹H NMR (DMSO-d₆300 MHz) δ 8.51 (d, J = 1.4, 1H), 8.47 (dd, J = 8.0, 1.4, 1H), 8.10-8.01 (m, 2H), 7.69-7.55 (m, 3H), 7.44-7.22 (m, 3H), 7.16 (d, J = 7.4, 1H), 4.08 (q, J = 6.6, 1H), 2.01 (s, 3H), 1.91 (s, 2H), 1.29 (d, J = 6.6, 3H). LC/MS (method B): 407.1 (M-NH₂)⁺. HPLC (method A) Rt 4.37min (purity: 99.8%).

Intermediates 45: N- Methyl radical -1-(3-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Methylamine hydrochloride

Step (ii) of 1: Methyl radical (3-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical ) Carbamic acid tert-butyl ester

A solution of intermediate 1 (2.01 g; 7.16 mmol; 1 eq.) in anhydrous THF (20 ml) was prepared in a round-bottom flask (50 ml) placed under a nitrogen atmosphere and cooled in an ice bath. N-methylmorpholine (0.83 ml; 7.52 mmol; 1.05 equiv) was added followed by isobutyl chloroformate (0.93 ml; 7.16 mmol; 1 equiv). The mixture was stirred in an ice bath for 1 hour. Intermediate 42 (2.0 g; 7.16 mmol; 1 eq.) was added in one portion and the cooling bath was removed. The reaction mixture was stirred at room temperature for 1 hour. It was then diluted with MTBE (40 ml) and washed with water (30 ml), 0.1N NaOH (20 ml), water (20 ml) and brine (20 ml). Drying (MgSO) ₄) The organic layer was concentrated under vacuum to give 3.97 g of a white foam. The foam was dissolved in toluene (30 ml) and the resulting solution was washed with waterThe solution was heated at 95 ℃ for 24 hours. The reaction mixture was concentrated in vacuo to give a yellow gum which was purified by flash chromatography (heptane/EtOAc gradient from 95:5 to 80: 20). The title compound was isolated as a colorless oil (3.25 g, 87%).¹H NMR (DMSO-d₆300 MHz) δ 8.53 (d, J = 1.4, 1H), 8.48 (dd, J = 8.0, 1.4, 1H), 8.15-7.96 (m, 2H), 7.66 (d, J = 8.0, 1H), 7.64-7.56 (m, 1H), 7.56-7.46 (m, 1H), 7.41-7.33 (m, 2H), 7.32-7.25 (m, 1H), 7.17 (d, J = 7.3, 1H), 4.49 (s, 2H), 2.81 (s, 3H), 2.02 (s, 3H), 1.47 (s, 9H). LC/MS (method B) 541.4 (M + NH)₄ ⁺). HPLC (method A) Rt 6.95 min. (purity: 98.7%).

Step (ii) of 2: N- Methyl radical -1-(3-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Methylamine hydrochloride

In a round-bottomed flask (100 ml), methyl (3- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) was prepared]-tert-butyl 1,2, 4-oxadiazol-3-yl } benzyl) carbamate (3.12 g; 5.96 mmol; 1 eq) in AcOH (18.72 ml) and then aqueous hydrochloric acid (2.93 ml of 32% aqueous solution; 29.80 mmol; 5.00 equivalents). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo to give a colorless oil. Et was added slowly ₂O (-40 ml) and the resulting precipitate was filtered off and treated with Et₂O (3X) then pentane (3X) and dried under vacuum (40 ℃ C., overnight). The title compound was isolated as a white powder (2.44 g, 89%).¹H NMR (DMSO-d₆, 300 MHz) δ9.34 (s, 2H), 8.55 (s, 1H), 8.50 (d, J = 8.0, 1H), 8.33 (s, 1H), 8.18 (d, J = 7.8, 1H), 7.83 (d, J = 7.8, 1H), 7.77 - 7.62 (m, 2H), 7.44 - 7.33 (m, 2H), 7.32 - 7.23 (m, 1H), 7.17 (d, J = 7.4, 1H)4.27 (s, 2H), 2.58 (s, 3H), 2.02 (s, 3H). LC/MS (method B) 424.3 (M + H)⁺. HPLC (method A) Rt 4.45min (purity: 99.7%). Elemental analysis: [ C ]₂₄H₂₀N₃OF₃- HCl]And (3) correction: 62.68% of C, 4.60% of H, 9.14% of N and 7.71% of Cl7; actually measuring: 62.56% of C, 4.76% of H, 8.81% of N and 7.45% of Cl7.

Intermediates 46: N-(1-{4-[(Z)- Amino group ( Oximino radical ) Methyl radical ] Phenyl radical }-2-{[ Tert-butyl radical ( Dimethyl group ) Silyl radical ] Oxy radical } Ethyl radical )-D- Alanine tert-butyl ester

Step (ii) of 1: 2- Hydroxy radical -1-(4- Cyanophenyl group ) Ethanones

To a solution of 2-bromo-1- (4-cyanophenyl) ethanone (15 g, 0.067 mol) in methanol (200 ml) was added sodium formate (13.6 g, 0.201 mol) at room temperature. The reaction mixture was refluxed for 15 h and filtered. The filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate, washed with water and brine solution, and taken up over Na₂SO₄Dried and evaporated. The crude product was purified by column chromatography using petroleum ether and ethyl acetate (60:40) as eluents to give (5.4 g, 50%) of the title compound as an off-white solid.¹H NMR: (DMSO-d₆, 400 MHz) δ8.06-8.04 (dd, J = 1.9, 6.7 Hz, 2H), 8.00-7.98 (dd, J = 1.9, 6.7 Hz, 2H), 5.27-5.24 (t, J = 5.9 Hz, 1H), 4.82-4.80 (d, J = 5.9 Hz, 2H)。

Step (ii) of 2: N-[1-(4- Cyanophenyl group )-2- Hydroxyethyl group ]-D- Alanine tert-butyl ester

To a solution of 2-hydroxy-1- (4-cyanophenyl) ethanone (3.2 g, 0.0198 mol) in methanol and DCM (1:1) (100 ml) was added D-alanine tert-butyl ester hydrochloride (3.6 g, 0.0198 mol) and triethylamine (6.9 ml, 0.0495 mol) at room temperature. The reaction mixture was heated at 70 ℃ for 18 h. The reaction mixture was cooled to 0 ℃ and NaBH was added portionwise₄(0.9 g, 0.0237 mol) and stirred at room temperature for 1 h. The reaction mixture was quenched with ice, extracted with DCM (2 × 100 ml), dried over sodium sulfate and evaporated. The crude product was purified by column chromatography using chloroform and methanol (90:10) as eluents to give (2.5 g, 43%) the title compound as a brown oil.¹H NMR: (CD₃OD, 400 MHz) δ7.71-7.69 (d, J = 8.36 Hz, 2H), 7.59-7.56 (d, J = 8.08 Hz, 2H), 4.76-4.74 (t, J = 6.32 Hz, 1H), 3.64-3.61 (q, 2H), 3.44-3.42 (d, J = 7.04Hz, 1H), 1.47 (s, 9H), 1.30-1.28 (d, J= 7.04Hz, 3H). LC/MS (method A): 291.3 (M + H)⁺. HPLC (method A) Rt 2.95 min. (purity: 76.8%).

Step (ii) of 3: N-[2-{[ Tert-butyl radical ( Dimethyl group ) Silyl radical ] Oxy radical }-1-(4- Cyanophenyl group ) Ethyl radical ]-D- Alanine tert-butyl ester

At room temperature, to N- [1- (4-cyanophenyl) -2-hydroxyethyl]To a solution of tert-butyl (2 g, 0.0068 mol) D-alanine in DCM (30 ml) was added imidazole (0.92 g, 0.0136 mol) and tert-butyldimethylsilyl chloride (1.25 g, 0.0082 mol). The reaction mixture was stirred at rt for 6 h and partitioned between DCM and water, the layers were separated and concentrated under reduced pressure. Using petroleum The crude product was purified by column chromatography using ether and ethyl acetate (80:20) as eluents to give (2 g, 74%) the title compound as a brown oil.¹H NMR: (DMSO-d₆, 400MHz) δ7.80-7.77 (m, 2H), 7.53-7.50 (m, 2H), 4.86-4.85 (t, J = 3.9 Hz, 1H ), 3.17-3.13 (m, 1H), 2.61-2.55 (m, 1H), 1.95-1.89 (m, 1H), 1.39 (s, 9H), 1.09-1.07 (d, J= 7.1 Hz, 3H), 0.80 (s, 9H), 0.04 (s,3H), -0.12 (s, 3H). LC/MS (method A): 405.3 (M + H)⁺. HPLC (method A) Rt 5.13 min. (purity: 73.7%).

Step (ii) of 4: N-(1-{4-[(Z)- Amino group ( Oximino radical ) Methyl radical ] Phenyl radical }-2-{[ Tert-butyl radical ( Dimethyl group ) Silyl radical ] Oxy radical } Ethyl radical )-D- Alanine tert-butyl ester

To N- [2- { [ tert-butyl (dimethyl) silyl group at room temperature]Oxy } -1- (4-cyanophenyl) ethyl]To a solution of tert-butyl (2 g, 0.0049 mol) of (D-alanine in) ethanol (40 ml) was added 50% aqueous hydroxylamine solution (1.63 ml, 0.0247 mol). The reaction mixture was heated at 70 ℃ for 6 h, cooled to room temperature, and concentrated under reduced pressure. The residue was dissolved in ethyl acetate, washed with water, brine and Na₂SO₄Dried and evaporated to give (2 g, 90%) the title compound as a brown oil.¹H NMR: (DMSO-d₆, 400MHz) δ9.57 (s, 1H), 7.61-7.59 (d, J = 8.3 Hz, 2H), 7.30-7.28 (d, J = 8.3 Hz, 2H), 5.76-5.74 (d, J= 5.00 Hz, 2H), 4.77-4.74 (m, 1H), 3.21-3.19 (m, 1H), 2.69-2.59 (m, 1H), 1.95-1.72 (m, 1H), 1.39 (s, 9H), 1.10-1.06 (m, 3H), 0.86-0.82 (m, 9H), 0.03 (s,3H), -0.12 (s, 3H). LC/MS (method A) 438.3 (M + H)⁺. HPLC (method A) Rt 3.82 min. (purity: 71.1%).

Examples 1: 2-[(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )( Methyl radical ) Amino group ] Butyric acid , Hydrochloride salt

Preparation of 2- [ (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 5 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) (methyl) amino]Tert-butyl butyrate. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.55 (brs, 1H), 8.51 (m, 1H), 8.07 (d, J =8.0 Hz, 1H), 8.00 (d, J =10.4 Hz, 1H), 7.93 (m, 1H), 7.68 (d, J =8 Hz, 1H), 7.43-7.33 (m, 2H), 7.29 (m, 1H), 7.17 (d, J = 7.5 Hz, 1H), 4.41 (m, 2H), 3.91 (m, 1H), 2.71 (brs, 3H), 2.02 (s, 3H), 1.97 (m, 2H), 1.00 (t, J =6.9 Hz, 3H). LC/MS (method B) 528.3 (M + H)⁺, 526.3 (M-H)^-. HPLC (method A) Rt 4.48 min (purity: 99.7%).

Examples 2: N-[(1R)-1-(4-{5-[2'-( Difluoromethyl group )-2- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2' - (difluoromethyl) -2-methylbiphenyl-4-yl) starting from intermediate 2 and intermediate 6 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white powder. ¹H NMR: (DMSO-d₆300 MHz) δ 8.17 (m, 3H), 8.08 (m, 1H), 7.76 (m, 3H), 7.65 (m, 2H), 7.44 (d, 1H, J =7.9 Hz), 7.33 (m, 1H), 6.64 (t, 1H, J = 54.6Hz), 4.50 (m, 1H), 3.51 (m, 1H), 2.14 (s, 3H), 1.59 (m, 3H), 1.40 (m, 3H). LC/MS (method B) 479.4 (M + H)⁺, 477.5 (M-H)^-. HPLC (method A) Rt 4.15 min (purity: 98.3%).

Examples 3: N-[(1S)-1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Preparation of N- [ (1S) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 10 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester. According to aIt was hydrolyzed as in operation 9 to give the title compound as a white foam.¹H NMR: (DMSO-d₆300 MHz) δ 8.33 (m, 1H), 8.18 (m, 3H), 7.80 (m, 2H), 7.43 (d, 1H, J =7.9 Hz), 7.39-7.23 (m, 3H), 7.14 (m, 1H), 4.53 (m, 1H), 4.20 (m, 2H), 3.66 (m, 1H), 3.25 (s, 3H), 2.03 (s, 3H), 1.62 (m, 3H), 1.44 (m, 3H). LC/MS (method B) 471.9 (M + H)⁺, 469.9 (M-H)^-. HPLC (method A) Rt 4.08 min (purity: 99.2%).

Examples 4: N-[(1R)-1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 6 according to general procedure 3 ]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white solid.¹H NMR: (DMSO-d₆300 MHz) δ 8.32 (m, 1H), 8.18 (m, 3H), 7.77 (m, 2H), 7.43 (d, 1H, J =7.9 Hz), 7.39-7.25 (m, 3H), 7.14 (m, 1H), 4.57 (m, 1H), 4.20 (m, 2H), 3.59 (m, 1H), 3.25 (s, 3H), 2.03 (s, 3H), 1.62 (m, 3H), 1.43 (m, 3H). LC/MS (method B): 472.2 (M + H)⁺, 470.2 (M-H)^-. HPLC (method A) Rt 4.07 min (purity: 99.3%).

Examples 5: N-[(1S)-1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [ (1S) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 8 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.32 (m, 1H), 8.18 (m, 3H), 7.78 (m, 2H), 7.44 (d, 1H, J =7.9 Hz), 7.39-7.25 (m, 3H), 7.14 (m, 1H), 4.51 (m, 1H), 4.20 (m, 2H), 3.84 (m, 1H), 3.59 (m, 1H), 3.25 (s, 3H), 2.03 (s, 3H), 1.64 (m, 3H). LC/MS (method B) 456.0 (M-H)^-. HPLC (method A) Rt 4.05 min (purity: 97.1%).

Examples 6: N-[(1R)-1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 6 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.18 (m, 3H), 8.06 (m, 1H), 7.91 (m, 1H), 7.83-7.65 (m, 4H), 7.14 (m, 2H), 4.52 (m, 1H), 3.51 (m, 1H), 2.11 (s, 3H), 1.60 (m, 3H), 1.41 (m, 3H). LC/MS (method B) 496.0 (M + H)⁺, 494.0 (M-H)^-. HPLC (method A) Rt 4.76 min (purity: 97.6%).

Examples 7: N-[(1R)-1-(4-{5-[2'-( Difluoromethyl group )-2- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2' - (difluoromethyl) -2-methylbiphenyl-4-yl) starting from intermediate 2 and intermediate 9 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆, 300 MHz) δ 8.19 (m, 3H), 8.09 (m, 1H), 7.78 (m, 3H), 7.65 (m, 2H), 7.44 (d, 1H, J=7.9 Hz), 7.33 (m, 1H), 6.64 (t, 1H, J= 54.6Hz), 4.51 (m, 1H), 3.84 (m, 1H), 3.59 (m, 1H), 2.14 (s3H), 1.64 (m, 3H). LC/MS (method B) 462.0 (M-H)^-. HPLC (method A) Rt 4.10 min (purity: 99.3%).

Examples 8: N-[(1R)-1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 9 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.18 (m, 3H), 8.08 (m, 1H), 7.91 (m, 1H), 7.78 (m, 3H), 7.69 (m, 1H), 7.42 (m, 2H), 4.51 (m, 1H), 3.83 (m, 1H), 3.58 (m, 1H), 2.12 (s, 3H), 1.63 (m, 3H). LC/MS (method B) 480.0 (M-H)^-. HPLC (method A) Rt 4.24 min (purity: 99.6%).

Examples 9: N-[(1R)-1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 9 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.32 (m, 1H), 8.18 (m, 3H), 7.79 (m, 2H), 7.43 (d, 1H, J =7.9 Hz), 7.38-7.26 (m, 3H), 7.14 (m, 1H), 4.51 (m, 1H), 4.20 (m, 2H), 3.81 (m, 1H), 3.57 (m, 1H), 3.25 (s, 3H), 2.03 (s, 3H), 1.64 (m, 3H). LC/MS (method B) 456.0 (M-H) ^-. HPLC (method A) Rt 3.87 min (purity: 95.1%).

Examples 10: (2S)-2-{[(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Amino group } Butyric acid , Hydrochloride salt

Following general procedure 3, starting from intermediate 1 and intermediate 12, preparation of (2S) -2- { [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Amino groupEthyl butyrate. It was hydrolyzed according to general procedure 9 to give the title compound as a white foam.¹H NMR: (DMSO-d₆300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.17 (m, 2H), 7.75 (m, 2H), 7.67 (m, 1H), 7.42-7.25 (m, 3H), 7.17 (m, 1H), 4.35 (m, 1H), 3.52 (m, 1H), 2.02 (s, 3H), 1.92-1.72 (m, 2H), 1.56 (m, 3H), 0.91 (t, 3H, J =7.4 Hz). LC/MS (method B) 510.0 (M + H)⁺, 508.0 (M-H)^-. HPLC (method A) Rt 4.42 min (purity: 97.5%).

Examples 11: (2R)-2-{[(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Amino group } Butyric acid , Hydrochloride salt

Starting from intermediate 1 and intermediate 13, according to general procedure 3, (2R) -2- { [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Amino } butanoic acid ethyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white powder. ¹H NMR: (DMSO-d₆300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.20 (m, 2H), 7.75 (m, 2H), 7.68 (m, 1H), 7.42-7.33 (m, 2H), 7.29 (m, 1H), 7.17 (m, 1H), 4.48 (m, 1H), 2.02 (s, 3H), 1.92-1.72 (m, 2H), 1.62 (m, 3H), 0.90 (t, 3H, J =7.4 Hz). LC/MS (method B) 510.0 (M + H)⁺, 508.0 (M-H)^-. HPLC (method A) Rt 4.43 min (purity: 99.3%).

Examples 12: N-(3- Fluorine -5-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical ) Leucine , Hydrochloride salt

Preparation of N- (3-fluoro-5- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 14 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) leucine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.55 (brs, 1H), 8.51 (m, 1H), 8.20 (m, 1H), 7.98 (m, 1H), 7.70 (m, 2H), 7.45-7.22 (m, 3H), 7.17 (m, 1H), 4.35 (m, 2H), 3.94 (m, 1H), 2.02 (s, 3H), 1.85-1.68 (m, 3H), 0.93 (m, 6H). LC/MS (method B): 542.3 (M + H)⁺, 540.3 (M-H)^-. HPLC (method A) Rt 5.29 min (purity: 98.9%).

Examples 13: N-(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-N- Methylvaline , Hydrochloride salt

Preparation of N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 15 according to general procedure 3 ]-1,2, 4-oxadiazol-3-yl } benzyl) -N-methylvaline tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (CD₃OD, 300 MHz) Δ 8.63 (brs, 1H), 8.50 (m, 1H), 8.15 (m, 1H), 8.08 (m, 1H), 7.81 (m, 1H), 7.58 (m, 1H), 7.39-7.20 (m, 3H), 7.14 (m, 1H), 4.54 (m, 2H), 4.01 (m, 1H), 2.89 (s, 3H), 2.58 (m, 1H), 2.06 (s, 3H), 1.24 (m, 3H), 1.09 (m, 3H). LC/MS (method B) 540.4 (M-H)^-. HPLC (method A) Rt 5.16 min (purity: 97%).

Examples 14: N- Methyl radical -N-[1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N-methyl-N- [1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 16 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to obtain the title compoundThe title compound was a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.19 (m, 3H), 8.07 (m, 1H), 7.91 (m, 1H), 7.87-7.75 (m, 3H), 7.69 (m, 1H), 7.42 (m, 2H), 4.78 (m, 1H), 4.02 (m, 2H), 2.80 (s, 3H), 2.11 (s, 3H), 1.71 (m, 3H). LC/MS (method B) 496.2 (M + H)⁺, 494.3 (M-H)^-. HPLC (method A) Rt 4.78 min (purity: 99%).

Examples 15: N-[1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-N- Methyl glycine , Hydrochloride salt

Preparation of N- [1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] starting from intermediate 3 and intermediate 16 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-N-methylglycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a colorless oil.¹H NMR: (DMSO-d₆300 MHz) delta 8.33 (brs, 1H), 8.19 (m, 3H), 7.82 (m, 2H), 7.43 (m, 1H), 7.39-7.25 (m, 3H), 7.14 (m, 1H), 4.74 (m, 1H), 4.19 (m, 2H), 3.98 (m, 2H), 3.25 (s, 3H), 2.76 (s, 3H), 2.04 (s, 3H), 1.69 (m, 3H). LC/MS (method B): 472.2 (M + H)⁺, 470.3 (M-H)^-. HPLC (method A) Rt 4.05 min (purity: 98.8%).

Examples 16: N-(2- Fluorine -4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-2- Methyl alanine , Hydrochloride salt

Preparation of N- (2-fluoro-4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 17 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.33 (brs, 1H), 8.18 (m, 1H), 8.08 (m, 1H), 7.98 (m, 1H), 7.89 (m, 1H), 7.44 (m, 1H), 7.39-7.25 (m, 3H), 7.14 (m, 1H), 4.28 (m, 2H), 4.19 (m, 2H), 3.25 (s, 3H), 2.04 (s, 3H), 1.61 (s, 6H). LC/MS (method B) 490.2 (M + H) ⁺, 488.3 (M-H)^-. HPLC (method A) Rt 4.08 min (purity: 98.7%).

Examples 17: N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Preparation of N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and methyl intermediate 10 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white foam.¹H NMR: (DMSO-d₆300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.21 (m, 2H), 7.83 (m, 2H), 7.67 (m, 1H), 7.36 (m, 2H), 7.29 (m, 1H), 7.17 (m, 1H), 4.54 (m, 1H), 3.67 (m, 1H), 2.02 (s, 3H), 1.63 (d, J = 6.8 Hz, 3H), 1.46 (d, J =7.1Hz, 3H). LC/MS (method B) 496.4 (M + H)⁺. HPLC (method A) Rt 4.85 min (purity: 98.4%).

Examples 18: N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine , Hydrochloride salt

Preparation of N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 11 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white foam. ¹H NMR: (DMSO-d₆, 300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.21 (m, 2H), 7.80 (m, 2H), 7.66 (m, 1H), 7.42-7.33 (m, 2H), 7.29 (m, 1H), 7.16 (m, 1H), 4.59 (m, 1H), 3.57 (m1H), 2.02 (s, 3H), 1.63 (d, J = 6.9 Hz, 3H), 1.45 (d, J =7.0Hz, 3H). LC/MS (method B) 496.4 (M + H)⁺. HPLC (method A) Rt 4.86 min (purity: 96.9%).

Examples 19: N-[(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 7 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white foam.¹H NMR: (DMSO-d₆300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.21 (m, 2H), 7.80 (m, 2H), 7.67 (m, 1H), 7.42-7.33 (m, 2H), 7.29 (m, 1H), 7.17 (m, 1H), 4.53 (m, 1H), 3.65 (m, 1H), 2.02 (s, 3H), 1.61 (d, J = 6.8 Hz, 3H), 1.44 (d, J =7.0Hz, 3H). LC/MS (method B) 496.5 (M + H)⁺, 494.5 (M-H)^-. HPLC (method A) Rt 4.89 min (purity: 100%).

Examples 20: N-[(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 6 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl ]-D-alanine methyl ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white solid.¹H NMR: (DMSO-d₆300 MHz) δ 8.55 (d, J = 1.5 Hz, 1H), 8.50 (dd, J = 8.0, 1.6 Hz, 1H), 8.21 (d, J = 8.5 Hz, 2H), 7.80 (d, J = 8.3 Hz, 2H), 7.68 (d, J = 8.1 Hz, 1H), 7.44-7.25 (m, 3H), 7.17 (d, J = 7.5 Hz, 1H), 4.59 (q, J = 6.8 Hz, 1H), 3.59 (q, J = 7.0 Hz, 1H), 2.02 (s, 3H), 1.64 (d, J = 6.8 Hz, 3H), 1.45 (d, J = 7.1 Hz, 3H). LC/MS (method B) 496.5 (M + H)⁺, 494.4 (M-H)^-. HPLC (method A) Rt 4.88 min (purity: 100%).

Examples 21: N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 8 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a pale yellow powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.21 (m, 2H), 7.79 (m, 2H), 7.69 (d, J = 7.9 Hz, 1H), 7.44-7.25 (m, 3H), 7.17 (m, 1H), 4.51 (m, 1H), 3.82 (m, 1H), 3.57 (m, 1H), 2.02 (s, 3H), 1.64 (d, J = 6.8 Hz, 3H). LC/MS (method B) 482.4 (M + H) ⁺, 480.4 (M-H)^-. HPLC (method A) Rt 4.28 min (purity: 96.4%).

Examples 22: N-[(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 9 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a pale yellow powder.¹H NMR: (DMSO-d₆, 300 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.21 (m, 2H), 7.79 (m, 2H), 7.69 (d, J = 7.9 Hz, 1H), 7.43-7.24 (m, 3H), 7.17 (m, 1H), 4.51 (m, 1H), 3.79 (m, 1H), 3.55(m, 1H), 2.02 (s, 3H), 1.63 (d, J = 6.8 Hz, 3H). LC/MS (method B) n.d (M + H)⁺, 480.5 (M-H)^-. HPLC (method A) Rt 4.30 min (purity: 99%).

Examples 23: N-[1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Step (ii) of 1: [1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Carbamic acid tert-butyl ester

The title compound was prepared following general procedure 3, starting from intermediate 4 and intermediate 18. It was isolated as a white foam. LC/MS (method B) 524.3 (M + H)⁺. HPLC (method A) Rt 6.31 min (purity: 99.6%).

Step (ii) of 2: 1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethylamine hydrochloride salt

According to general procedure 8, starting from [1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) ]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]The title compound was prepared starting from tert-butyl carbamate and isolated as a white foam (620 mg, quantitative).¹H NMR (DMSO-d₆300 MHz) δ 8.60 (brs, 3H), 8.18 (m, 3H), 8.07 (m, 1H), 7.91 (d, J =8 Hz, 1H), 7.77 (m, 3H), 7.68 (m, 1H), 7.41 (m, 1H), 4.53 (m, 1H), 2.11 (s, 3H), 1.56 (d, J =6.8 Hz, 3H). LC/MS (method B) n.dp (M + H), n.dp (M-H). HPLC (method A) Rt 4.22 min (purity: 99.7%).

Step (ii) of 3: N-[1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

According to general procedure 11, starting from 1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl)]Starting from (E) -1,2, 4-oxadiazol-3-yl } phenyl) ethylamine hydrochloride and tert-butyl bromoacetate, N- [1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] was prepared]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.18 (m, 3H), 8.08 (m, 1H), 7.91 (d, J =7.90Hz, 1H), 7.77 (m, 3H), 7.69 (m, 1H), 7.41 (m, 2H), 4.74 (m, 1H), 3.74 (m, 1H), 3.52 (m, 1H), 2.11 (s, 3H), 1.61 (d, J =6.8 Hz, 3H). LC/MS (method B): 482.2 (M + H) ⁺, 480.2 (M-H)^-. HPLC (method A) Rt 4.25 min (purity: 99.9%).

Examples 24: N-[(1S)-1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine , Hydrochloride salt and N-[(1R)-1-(4-{5-[2- methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Methyl N- [1- (4- {5- [ 2-methyl-2 '- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -alanine was prepared according to general procedure 11 starting from 1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethylamine hydrochloride (example 23, step 2) and methyl 2-bromopropionate. The resulting 2 diastereomeric pairs were separated by flash chromatography (heptane/EtOAc gradient from 95:5 to 85: 15) to give methyl N- [ (1S) -1- (4- {5- [ 2-methyl-2 '- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alaninate and methyl N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alaninate (75 mg, 27%) as the first fractions, and as a second fraction methyl N- [ (1S) -1- (4- {5- [ 2-methyl-2 '- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine and methyl N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine (50 mg, 18%).

According to general procedure 9, a solution containing N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) was hydrolyzed]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester and N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-a second fraction of methyl D-alanine to give the title compound as a white powder.¹H NMR: (DMSO-d₆400 MHz) δ 8.15 (m, 3H), 8.07 (m, 1H), 7.91 (m, 1H), 7.83-7.64 (m, 4H), 7.41 (m, 2H), 4.35 (m, 1H), 2.11 (s, 3H), 1.53 (d, J = 6.5 Hz, 3H), 1.35 (d, J =7.0 Hz, 3H). LC/MS (method B) 496.2 (M + H)⁺, 494.3 (M-H)^-. HPLC (method A) Rt 4.28 min (purity: 100%). N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) can be isolated by chiral HPLC]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester and N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester.

Examples 25: N-[(1S)-1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine and N-[(1R)-1-(4-{5-[2- methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine

According to the general procedure 9, N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) was hydrolyzed ]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester and N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester (prepared in example 24) to give the title compound as a white powder.¹H NMR: (DMSO-d₆400 MHz) δ 8.15 (m, 3H), 8.07 (m, 1H), 7.91 (m, 1H), 7.79 (m, 1H), 7.69 (m, 3H), 7.41 (m, 2H), 4.34 (m, 1H), 2.11 (s, 3H), 1.51 (d, J = 6.6 Hz, 3H), 1.32 (d, J =7.1 Hz, 3H). LC/MS (method B) 496.2 (M + H)⁺, 494.3 (M-H)^-. HPLC (method A) Rt 4.29 min (purity: 86.5%). N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) can be isolated by chiral HPLC]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine ester and N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl)]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester.

Examples 26: N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine , Hydrochloride salt and N-[(1R)-1-(4-{5-[2'- methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Step (ii) of 1: [1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Carbamic acid tert-butyl ester

The title compound was prepared following general procedure 3 starting from intermediate 1 and intermediate 18 and isolated as a white foam. LC/MS (method B) 524.4 (M + H) ⁺, 522.4 (M-H)^-. HPLC (method A) Rt 6.43 min (purity: 99.6%).

Step (ii) of 2: 1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethylamine, hydrochloride salt.

Following general procedure 8, starting from [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl)]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]The title compound was prepared starting from tert-butyl carbamate and isolated as a yellow foam (880 mg, 91%).¹H NMR: (DMSO-d₆, 400 MHz) δ 8.58-8.47 (m, 4H), 8.20 (d, J =8.3 Hz, 2H), 7.76 (d, J =8.3 Hz, 2H), 7.67 (d, J = 8.0 Hz, 1H), 7.42-7.25 (m, 3H), 7.17 (d, J = 7.3 Hz, 1H), 4.54 (m, 1H), 2.02 (s, 3H), 1.56 (d, J =6.8 Hz, 3H). LC/MS (method B) 459.2 (M-H)^-. HPLC (method A) Rt 4.32 min (purity: 98.7%).

Step (ii) of 3: N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine , Hydrochloride salt and N-[(1R)-1-(4-{5-[2'- methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Methyl N- [1- (4- {5- [2 '-methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -alanine was prepared according to general procedure 11 starting from 1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethylamine hydrochloride and methyl 2-bromopropionate. The resulting 2 diastereomeric pairs can be separated by flash chromatography (heptane/EtOAc gradient from 95:5 to 85: 15) to yield as first fractions methyl N- [ (1S) -1- (4- {5- [2 '-methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine and methyl N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine (90 mg, 33%), and as a second fraction methyl N- [ (1S) -1- (4- {5- [2 '-methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -L-alanine and methyl N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine (85 mg, 31%).

Hydrolysis of a mixture containing N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) according to general procedure 9]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester and N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-a second fraction of methyl D-alanine to give the title compound as a white powder.¹H NMR: (DMSO-d₆400 MHz) δ 8.54 (brs, 1H), 8.49 (m, 1H), 8.19 (d, J =8.3Hz, 2H), 7.79 (d, J =8.3Hz, 2H), 7.67 (d, J =8.3Hz, 1H), 7.42-7.25 (m, 3H), 7.16 (m, 1H), 4.44 (m, 1H), 3.50 (m, 1H), 2.02 (s, 3H), 1.58 (d, J = 6.6 Hz, 3H), 1.40 (d, J =7.0 Hz, 3H). LC/MS (method B) 496.2 (M + H)⁺, 494.3 (M-H)^-. HPLC (method A) Rt 4.36 min (purity: 97.8%). N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] can be isolated by chiral HPLC according to, for example, method J described above]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine ester and N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester.

Examples 27: N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt and N-[(1R)-1-(4-{5-[2'- methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-L- Alanine , Hydrochloride salt

N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) was hydrolyzed according to the general procedure 9]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine methyl ester and N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester (prepared in example 26) to give the title compound as a white powder.¹H NMR: (DMSO-d₆400 MHz) δ 8.54 (brs, 1H), 8.50 (m, 1H), 8.20 (d, J =8.3Hz, 2H), 7.75 (d, J =8.3Hz, 2H), 7.67 (d, J =8.0Hz, 1H), 7.42-7.25 (m, 3H), 7.16 (m, 1H), 4.53 (m, 1H), 3.55 (m, 1H), 2.02 (s, 3H), 1.60 (d, J = 6.6 Hz, 3H), 1.41 (d, J =7.0 Hz, 3H). LC/MS (method B) 496.2 (M + H)⁺, 494.3 (M-H)^-. HPLC (method A) Rt 4.34 min (purity: 91.1%). N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] can be isolated by chiral HPLC according to, for example, method J described above]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-D-alanine ester and N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-L-alanine methyl ester.

Examples 28: N- Methyl radical -N-[1-(3-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Alanine , Hydrochloride salt

Preparation of N-methyl-N- [1- (3- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 19 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]An alanine ester. It was hydrolyzed according to general procedure 9 to give the title compound as a colorless oil.¹H NMR: (DMSO-d₆300 MHz) δ 8.27 (m, 1H), 8.17 (brs, 1H), 8.09 (m, 2H), 7.91 (d, J =8Hz, 1H), 7.78 (m, 2H), 7.68 (m, 2H), 7.41 (m, 2H), 4.48 (m, 1H), 3.81 (m, 1H), 2.54 (m, 3H), 2.11 (s, 3H), 1.58 (m, 3H), 1.37 (m, 3H). LC/MS (method B) 510.3 (M + H)⁺, 508.4 (M-H)^-. HPLC (method A) Rt 4.33 min (purity: 95.1%).

Examples 29: N- Methyl radical -N-[1-(3-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Alanine , Hydrochloride salt

Preparation of N-methyl-N- [1- (3- {5- [2' -methyl-2 ] starting from intermediate 1 and intermediate 19 according to general procedure 3- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]An alanine ester. It was hydrolyzed according to general procedure 9 to give the title compound as a white foam.¹H NMR: (DMSO-d₆300 MHz) delta 8.54 (brs, 1H), 8.51 (m, 1H), 8.30 (m, 1H), 8.14 (m, 1H), 7.80 (m, 1H), 7.68 (m, 2H), 7.43-7.33 (m, 2H), 7.29 (m, 1H), 7.17 (m, 1H), 4.51 (m, 1H), 3.88 (m, 1H), 2.57 (m, 3H), 2.02 (s, 3H), 1.59 (m, 3H), 1.39 (m, 3H). LC/MS (method B) 510.3 (M + H) ⁺, 508.4 (M-H)^-. HPLC (method A) Rt 4.41 min (purity: 96.1%).

Examples 30: N- Methyl radical -N-[1-(3-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N-methyl-N- [1- (3- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 20 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.33 (m, 1H), 8.20 (m, 2H), 8.08 (m, 1H), 7.92 (m, 1H), 7.85 (m, 1H), 7.81-7.65 (m, 3H), 7.42 (m, 2H), 4.76 (m, 1H), 3.95 (m, 2H), 2.77 (s, 3H), 2.12 (s, 3H), 1.71 (m, 3H). LC/MS (method B) 496.2 (M + H)⁺, 494.3 (M-H)^-. HPLC (method A) Rt 4.28 min: (method A) ((method A))Purity: 98.6%).

Examples 31: N-(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-N,2- Dimethyl alanine , Hydrochloride salt

Preparation of N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 21 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -N, 2-dimethylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder. ¹H NMR: (DMSO-d₆300 MHz) δ 8.56 (m, 1H), 8.52 (m, 1H), 8.11 (m, 1H), 8.03 (m, 1H), 7.93 (m, 1H), 7.69 (d, J =8Hz, 1H), 7.43-7.33 (m, 2H), 7.29 (m, 1H), 7.17 (m, 1H), 3.48 (m, 2H), 2.66 (brs, 3H), 2.03 (s, 3H), 1.61 (brs, 6H). LC/MS (method B) 528.1 (M + H)⁺, 526.3 (M-H)^-. HPLC (method A) Rt 4.45 min (purity: 97.6%).

Examples 32: N-[1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 22 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.55 (m, 1H), 8.51 (m, 1H), 8.22 (d, J =8.4 Hz, 2H), 7.77 (d, J =8.4 Hz, 2H), 7.68 (d, J =8Hz, 1H), 7.43-7.25 (m, 3H), 7.17 (m, 1H), 4.51 (m, 1H), 3.85 (m, 1H), 3.59 (m, 1H), 2.02 (s, 3H), 1.63 (d, J =6.8 Hz, 3H). LC/MS (method B) 482.0 (M + H)⁺, 480.1 (M-H)^-. HPLC (method A) Rt 4.82 min (purity: 99.7%).

Examples 33: N- Methyl radical -N-[(1S)-1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N-methyl-N- [ (1S) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4) starting from intermediate 4 and intermediate 23 according to general procedure 4-radical]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. Following general procedure 8, it was deprotected to give the title compound as a white powder. Melting point: 217 deg.C.-28.2 (c 1.47, EtOH)。¹H NMR (DMSO-d₆300 MHz) δ 8.25-8.14 (m, 3H), 8.07 (dd, J = 8.0, 1.4 Hz, 1H), 7.91 (d, J = 7.5 Hz, 1H), 7.86-7.74 (m, 3H), 7.68 (t, J = 7.6 Hz, 1H), 7.46-7.35 (m, 2H), 4.76 (q, J = 6.9 Hz, 1H), 4.11-3.89 (m, 2H), 2.79 (s, 3H), 2.11 (s, 3H), 1.71 (d, J = 7.0 Hz, 3H). LC/MS (method B) 494.3 (M-H)^-, 496.2 (M+H)⁺. HPLC (method A) Rt 4.78 min (purity: 99.7%). Elemental analysis: [ C ]₂₇H₂₄N₃O₃F₃ - HCl-0.2 H₂O]And (3) correction: c60.55%, H4.78%, N7.85%, Cl 6.62%; actually measuring: c60.41%, H4.83%, N7.91%, Cl 6.71%.

Examples 34: N- Methyl radical -N-[(1S)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N-methyl-N- [ (1S) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 23 according to general procedure 4]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. Following general procedure 8, it was deprotected to give the title compound as a white powder. Melting point: 221 ℃. -27.4 (c 1.54, EtOH)。¹H NMR (DMSO-d₆300 MHz) δ 8.55 (d, J = 1.5 Hz, 1H), 8.50 (dd, J = 8.0, 1.5 Hz, 1H), 8.23 (d, J = 8.6 Hz, 2H), 7.82 (d, J = 8.3 Hz, 2H), 7.66 (d, J = 8.1 Hz, 1H), 7.41-7.25 (m, 3H), 7.16 (d, J = 7.5 Hz, 1H), 4.75 (q, J = 6.9 Hz, 1H), 4.10-3.88 (m, 2H), 2.78 (s, 3H), 2.01 (s, 3H), 1.70 (d, J = 6.9 Hz, 3H). LC/MS (method A) 494.3 (M-H)^-, 496.2 (M+H)⁺. HPLC (method A) Rt 4.85 min (purity: 99.8%). Elemental analysis: [ C ]₂₇H₂₄N₃O₃F₃- HCl-0.2 H₂O]And (3) correction: c60.55%, H4.78%, N7.85%, Cl 6.62%; actually measuring: c60.52%, H4.63%, N7.90%, Cl 6.65%.

Examples 35: N- Methyl radical -N-[(1R)-1-(4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Following general procedure 4, starting from intermediate 4 and intermediate 24, N-methyl-N- [ (1R) -1- (4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl was prepared]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. In accordance with the general procedure 8,it was deprotected to give the title compound as a white powder. Melting point: 217 deg.C. 28.1 (c 1.41, EtOH)。¹H NMR (DMSO-d₆300 MHz) δ 8.25-8.14 (m, 3H), 8.07 (dd, J = 8.0, 1.4 Hz, 1H), 7.91 (d, J = 7.5 Hz, 1H), 7.86-7.74 (m, 3H), 7.68 (t, J = 7.6 Hz, 1H), 7.46-7.35 (m, 2H), 4.76 (q, J = 6.9 Hz, 1H), 4.11-3.89 (m, 2H), 2.79 (s, 3H), 2.11 (s, 3H), 1.71 (d, J = 7.0 Hz, 3H). LC/MS (method B) 494.3 (M-H) ^-, 496.2 (M+H)⁺. HPLC (method A) Rt 4.79 min (purity: 99.8%). Elemental analysis: c₂₇H₂₄N₃O₃F₃- HCl-0.2 H₂O]And (3) correction: c60.55%, H4.78%, N7.85%, Cl 6.62%; actually measuring: c60.44%, H4.64%, N7.89%, Cl 6.67%.

Examples 36: N- Methyl radical -N-[(1R)-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N-methyl-N- [ (1R) -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 24 according to general procedure 4]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. Following general procedure 8, it was deprotected to give the title compound as a white powder. Melting Point：221℃。 26.1 (c 1.44, EtOH)。¹H NMR (DMSO-d₆300 MHz) δ 8.55 (d, J = 1.5 Hz, 1H), 8.50 (dd, J = 8.0, 1.5 Hz, 1H), 8.23 (d, J = 8.6 Hz, 2H), 7.82 (d, J = 8.3 Hz, 2H), 7.66 (d, J = 8.1 Hz, 1H), 7.41-7.25 (m, 3H), 7.16 (d, J = 7.5 Hz, 1H), 4.75 (q, J = 6.9 Hz, 1H), 4.10-3.88 (m, 2H), 2.78 (s, 3H), 2.01 (s, 3H), 1.70 (d, J = 6.9 Hz, 3H). LC/MS (method B) 494.3 (M-H)^-, 496.2 (M+H)⁺. HPLC (method A) Rt 4.87 min (purity: 99.8%). Elemental analysis [ C₂₇H₂₄N₃O₃F₃- HCl- 0.2 H₂O]And (3) correction: c60.55%, H4.78%, N7.85%, Cl 6.62%; actually measuring: c60.37%, H4.79%, N7.84%, Cl 6.65%.

Examples 37: N-[1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-N- Methyl alanine

Preparation of N- [1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl ] starting from intermediate 3 and intermediate 25 according to general procedure 4]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-methyl N-methylalaninate. It was deprotected according to general procedure 9 to give the title compound as a white powder. LC/MS (method B) 484.4 (M-H)^-, 486.3 (M+H)⁺. HPLC (method A) Rt 4.62 min (purity: 98.3%).

Examples 38: N-[1-(3-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [1- (3- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 26 according to general procedure 4]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. Following general procedure 8, it was deprotected to give the title compound as a pale yellow powder. Melting point: 176 deg.c.¹H NMR (DMSO-d₆300 MHz) δ 8.33 (s, 1H), 8.20-8.13 (m, 2H), 8.08 (dd, J = 8.0, 1.5 Hz, 1H), 7.92 (d, J = 7.7 Hz, 1H), 7.80 (t, J = 7.4 Hz, 2H), 7.70 (td, J = 7.7, 2.6 Hz, 2H), 7.42 (t, J = 8.6 Hz, 2H), 4.54 (q, J = 6.7 Hz, 1H), 3.83 (d, J = 17.0 Hz, 1H), 3.56 (d, J = 16.8 Hz, 1H), 2.12 (s, 3H), 1.65 (d, J = 6.8 Hz, 3H). LC/MS (method A) 484.5 (M-H) ^-, 486.4 (M+H)⁺. HPLC (method A) Rt 4.76 min (purity: 96.3%).

Examples 39: N- Methyl radical -N-[1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Alanine

Step (ii) of 1: 4-(1- Hydroxyethyl group ) Benzonitrile

4-Acetylbenzonitrile (2.0 g; 13.8 mmol) was dissolved in THF (10 mL) and MeOH (10 mL). Sodium borohydride (782 mg; 20.7 mmol) was added portionwise at 0 deg.C and the reaction stirred at room temperature for 40 min. The solvent was removed under vacuum, EtOAC was added, and H was used₂O washing the organic phase over MgSO₄Dried, filtered, and concentrated to give the title compound as a colorless oil (2.3 g, quantitative). HPLC (method A) Rt 4.31 min (purity: 99.2%).

Step (ii) of 2: N'- Hydroxy radical -4-(1- Hydroxyethyl group ) Benzamidine

The title compound was prepared according to general procedure 1 starting from 4- (1-hydroxyethyl) benzonitrile (2.3 g; 16.8 mmol) obtained in step 1 to give 2.5 g (88%) as a colorless foam. LC/MS (method B) 181.0 (M + H)⁺。

Step (ii) of 3: 1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethanol

Following general procedure 3, N' -hydroxy-4 obtained from intermediate 1 and in step 2Starting with (1-hydroxyethyl) benzamidine, 1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethanol to give the title compound as a pale yellow oil (1.9 g, 80%). ¹H NMR (DMSO-d₆300 MHz) δ 8.64 (d, J = 1.3 Hz, 1H), 8.40 (dd, J = 8.0, 1.4 Hz, 1H), 8.19 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.1 Hz, 2H), 7.47 (d, J = 8.0 Hz, 1H), 7.38-7.21 (m, 3H), 7.16 (d, J = 7.5 Hz, 1H), 5.01 (q, J = 6.5 Hz, 2H), 2.07 (s, 3H), 1.56 (d, J = 6.6 Hz, 3H). LC/MS (method B) 425.1 (M + H)⁺. HPLC (method A) Rt 5.60 min (purity: 100.0%).

Step (ii) of 4: 1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethanones

To a stirred solution of oxalyl chloride (758 μ L; 8.8 mmol) in anhydrous DCM (50 mL) was added a solution of dimethyl sulfoxide (1.3 mL; 17.7 mmol) dissolved in DCM (10 mL) at-68 ℃ over 5 min. The reaction mixture was stirred for 15 min, followed by addition of 1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) over 15 min]-1,2, 4-oxadiazol-3-yl } phenyl) ethanol (1.5 g; 3.5 mmol, obtained in step 3) in DCM (10 mL). After stirring at-68 ℃ for 1h, the reaction was allowed to reach-30 ℃ and stirred for 15 min, then cooled again to-68 ℃. Triethylamine (1.8 mL; 12.7 mmol) was added and the mixture was allowed to reach room temperature. The clear yellow solution was partitioned between saturated aqueous sodium bicarbonate and ethyl acetate, the organic layer was washed with brine, over MgSO ₄Dried, filtered, and concentrated. The crude product mixture was purified by flash chromatography to give the title compound as a white powder.¹H NMR (DMSO-d₆, 300 MHz) δ8.67-8.47 (m, 2H), 8.29 (d, J = 7.9 Hz, 2H), 8.19 (d, J = 8.3 Hz, 2H), 7.67 (d, J = 7.9 Hz, 1H), 7.43-7.25 (m, 3H) 7.17 (d, J = 7.7 Hz, 1H), 2.67 (s, 3H), 2.02 (s, 3H). HPLC (method A) Rt 6.00 min (purity: 99.0%).

Step (ii) of 5: N- Methyl radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethylamine (ethylamine)

To the 1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl group obtained in step 4]-1,2, 4-oxadiazol-3-yl } phenyl) ethanone (515 mg; 1.2 mmol) in THF (5 mL), methylamine (732 μ L; 2.00M; 1.5 mmol) and titanium isopropoxide (199 μ L; 0.67 mmol). The mixture was stirred at room temperature overnight. Sodium borohydride (194 mg; 5.1 mmol) was added and the reaction stirred at room temperature for 2 h. Then 20 mL NH was added₄OH, and the reaction was stirred at room temperature for 1 hour 30 minutes. Then filtered through a celite pad, washing the pad with EtOAc. With saturated NaHCO₃The organic phase was washed with brine over MgSO₄After drying, filtration and concentration, and purification by flash chromatography, the title compound was obtained as a colorless oil.¹H NMR (DMSO-d₆300 MHz) δ 8.66 (s, 1H), 8.43 (d, J = 7.6 Hz, 1H), 8.27 (d, J = 8.1 Hz, 2H), 7.49 (t, J = 8.8 Hz, 3H), 7.42-7.23 (m, 3H), 7.17 (d, J = 7.4 Hz, 1H), 3.94-3.80 (m, 1H), 3.61 (br s, 1H), 2.44 (d, J = 5.7 Hz, 3H), 2.09 (s, 3H), 1.77 (d, J = 6.8 Hz, 3H). LC/MS (method B) 438.2 (M + H) ⁺. HPLC (method A) Rt 6.07 min (purity: 78.6%).

Step (ii) of 6: N- Methyl radical -N-[1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Alanine methyl ester

Following general procedure 10, from N-methyl-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) obtained in step 5]Starting from-1, 2, 4-oxadiazol-3-yl } phenyl) ethanamine (40 mg, 0.09 mmol), the title compound is prepared to yield 46 mg (96%) as a colourless oil. LC/MS (method B) 524.3 (M + H)⁺. HPLC (method A) Rt 4.61 min (purity: 70.5%).

Step (ii) of 7: N- Methyl radical -N-[1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Alanine

According to general procedure 9, from N-methyl-N- [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] obtained in step 6]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Alanine methyl ester (40 mg; 0.08 mmol) the title compound was prepared. After purification using mass-directed Autoprep, it was isolated as a white solid. LC/MS (method B) 508.3 (M-H)^-, 510.2 (M+H)⁺. HPLC (method A) Rt 4.92 min (purity: 99.8%).

Examples 40: N- Methyl radical -N-[1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N-methyl-N- [1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 16 according to general procedure 4 ]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. Following general procedure 8, it was deprotected to give the title compound as a white powder. Melting point: 204 ℃. LC/MS (method B): 494.0 (M-H)^-, 496.0 (M+H)⁺. HPLC (method A) Rt 4.36 min (purity: 98.1%).

Examples 41: 2-[(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical ) Amino group ] Butyric acid , Hydrochloride salt

Preparation of 2- [ (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 27 according to general procedure 4]-1,2, 4-oxadiazol-3-yl } benzyl) amino]Tert-butyl butyrate. Following general procedure 8, it was deprotected to give the title compound as a white powder. Melting point: 214 deg.C.¹H NMR (DMSO-d₆300 MHz) δ 8.56 (d, J = 1.6 Hz, 1H), 8.51 (dd, J = 7.9, 1.5 Hz, 1H), 8.08 (dd, J = 7.8, 1.5 Hz, 1H), 7.99 (dd, J = 10.2, 1.5 Hz, 1H), 7.91 (t, J = 7.8 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.43-7.25 (m, 3H), 7.17 (d, J = 7.3 Hz, 1H), 4.30 (m, 2H), 4.06-3.96 (m, 1H), 2.08-1.87 (m, 5H), 0.97 (t, J = 7.4 Hz, 3H). LC/MS (method B) 512.2 (M-H)^-, 514.2 (M+H)⁺. HPLC (method A) Rt 4.88 min (purity: 99.7%).

Examples 42: N-[1-(3-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine , Hydrochloride salt

Preparation of N- [1- (3- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 26 according to general procedure 4]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]Glycine tert-butyl ester. Following general procedure 8, it was deprotected to give the title compound as a white powder. Melting point: 177 ℃.¹H NMR (DMSO-d₆300 MHz) δ 9.76 (br s, 1H), 8.56 (d, J = 1.3 Hz, 1H), 8.51 (dd, J = 7.9, 1.7 Hz, 1H), 8.36 (s, 1H), 8.19 (d, J = 8.0 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.75-7.65 (m, 2H), 7.42-7.25 (m, 3H), 7.17 (d, J = 7.3 Hz, 1H), 4.58 (q, J = 6.5 Hz, 1H), 3.89 (d, J = 16.8 Hz, 1H), 3.60 (d, J = 17.2 Hz, 1H), 2.03 (s, 3H), 1.66 (d, J = 6.8, 3H). LC/MS (method B) 480.2 (M-H)^-, 482.1 (M+H)⁺. HPLC (method A)Rt 4.83 min (purity: 99.4%).

Examples 43: N-(2- Fluorine -4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-N,2- Dimethyl alanine , Hydrochloride salt

Preparation of N- (2-fluoro-4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 29 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -N, 2-dimethylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder. ¹H NMR: (DMSO-d₆300 MHz) δ 8.34 (m, 1H), 8.18 (dd, J =8.0 Hz, 1H), 8.09 (dd, J =8.0 Hz, 1H), 8.03-7.91 (m, 2H), 7.45 (d, J =8Hz, 1H), 7.40-7.26 (m, 3H), 7.15 (d, J = 7.3 Hz, 1H), 4.38 (brs, 2H), 4.20 (m, 2H), 3.25 (s, 3H), 2.69 (s, 3H), 2.04 (s, 3H), 1.65 (s, 6H). LC/MS (method B) 504.3 (M + H)⁺, 502.4 (M-H)^-. HPLC (method A) Rt 4.16 min (purity: 99.8%).

Examples 44: N-(2- Fluorine -4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-N- Methyl alanine , Hydrochloride salt

Preparation of N- (2-fluoro-4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 3 and intermediate 30 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -N-methylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.33 (m, 1H), 8.17 (dd, J =8.0 Hz, 1H), 8.06 (dd, J =8.0 Hz, 1H), 8.02-7.93 (m, 2H), 7.43 (d, J =8Hz, 1H), 7.39-7.25 (m, 3H), 7.14 (d, J = 7.3 Hz, 1H), 4.47 (m, 2H), 4.33-4.12 (m, 2H), 3.25 (s, 3H), 2.76 (s, 3H), 2.03 (s, 3H), 1.59 (d, J = 7.2 Hz, 3H). LC/MS (method B) 504.3 (M + H)⁺, 502.4 (M-H)^-. HPLC (method A) Rt 4.10 min (purity: 99.5%).

Examples 45: N-(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-N- Methylserine , A hydrochloride salt.

Starting from intermediate 1 and intermediate 31, O- (tert-butyl) -N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) di-tert-butyl) according to general procedure 3Phenyl-4-yl]-1,2, 4-oxadiazol-3-yl } benzyl) -N-methylserine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.55 (brs, 1H), 8.51 (m, 1H), 8.06 (m, 1H), 7.97 (d, J =10.2 Hz, 1H), 7.91 (t, J = 7.7 Hz, 1H), 7.68 (d, J =8 Hz, 1H), 7.37 (m, 2H), 7.29 (m, 1H), 7.17 (d, J = 7.7 Hz, 1H), 4.41 (m, 2H), 4.40 (m, 3H), 2.73 (s, 3H), 2.03 (s, 3H). LC/MS (method B) 530.2 (M + H)⁺, 528.3 (M-H)^-. HPLC (method A) Rt 4.27 min (purity: 100%).

Examples 46: N-(3- Fluorine -5-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-2- Methyl alanine , A hydrochloride salt.

Preparation of N- (3-fluoro-5- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 32 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder. ¹H NMR: (DMSO-d₆300 MHz) δ 8.51 (brs, 1H), 8.47 (m, 1H), 8.15 (m, 1H), 7.88 (m, 1H), 7.67 (m, 2H), 7.36 (m, 2H), 7.28 (m, 1H), 7.16 (d, J = 7.6 Hz, 1H), 4.20 (brs, 2H), 2.02 (s, 3H), 1.51 (s, 6H). LC/MS (method B) 514.3 (M + H), 512.3 (M-H). HPLC (method A) Rt 4.44 min (purity: 99.2%).

Examples 47: N-(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical ) Valine , A hydrochloride salt.

Preparation of N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 33 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) valine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.55 (brs, 1H), 8.50 (m, 1H), 8.07 (m, 1H), 7.96 (m, 2H), 7.68 (d, J =8.2 Hz, 2H), 7.37 (m, 2H), 7.29 (m, 1H), 7.17 (d, J = 7.6 Hz, 1H), 4.30 (m, 2H), 3.90 (m, 1H), 2.41 (m, 1H), 2.02 (s, 3H), 1.08 (d, J =7 Hz, 3H), 097 (d, J =6.8 Hz, 3H). LC/MS (method B) 528.3 (M + H)⁺, 526.4 (M-H)^-. HPLC (method A) Rt 4.48 min (purity: 100%).

Examples 48: N-(2- Fluorine -4-{5-[2- Methyl radical -2'-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-2- Methyl alanine , A hydrochloride salt.

Preparation of N- (2-fluoro-4- {5- [ 2-methyl-2' - (trifluoromethyl) biphenyl-4-yl) starting from intermediate 4 and intermediate 34 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) delta 8.17 (brs, 1H), 8.06 (m, 2H), 8.01-7.89 (m, 3H), 7.79 (m, 1H), 7.68 (m, 1H), 7.72 (m, 2H), 4.27 (brs, 2H), 2.11 (s, 3H), 1.63 (s, 6H). LC/MS (method B): 514.3 (M + H)⁺, 512.4 (M-H)^-. HPLC (method A) Rt 4.31 min (purity: 99.7%).

Examples 49: N-(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-2- Methyl alanine , A hydrochloride salt.

Preparation of N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 34 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -2-methylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 9.87 (m, 1H), 8.55 (brs, 1H), 8.50 (d, J =8.1 Hz, 1H), 8.07 (m, 1H), 7.99 (m, 2H), 7.68 (d, J =8 Hz, 1H), 7.37 (m, 2H), 7.29 (m, 1H), 7.16 (m, 1H), 4.27 (brs, 2H), 2.02 (s, 3H), 1.63 (s, 6H). LC/MS (method B) 514.4 (M + H) ⁺, 512.4 (M-H)^-. HPLC (method A) Rt 4.37 min (purity: 100%).

Examples 50: N-(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )-N- Methyl alanine , A hydrochloride salt.

Preparation of N- (2-fluoro-4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) starting from intermediate 1 and intermediate 30 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } benzyl) -N-methylalanine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆300 MHz) δ 8.55 (brs, 1H), 8.51 (m, 1H), 8.09 (m, 1H), 7.99 (m, 2H), 7.68 (d, J =8.1 Hz, 1H), 7.37 (m, 2H), 7.29 (m, 1H), 7.17 (d, J = 7.5 Hz, 1H), 4.47 (m, 2H), 4.26 (brs, 1H), 2.75 (s, 3H), 2.02 (s, 3H), 1.59 (d, J =7.1 Hz, 3H). LC/MS (method B) 514.4 (M + H)⁺, 512.4 (M-H)^-. HPLC (method A) Rt 4.40 min (purity: 100%).

Examples 51: 2-((2- Methoxy radical -1-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Ethyl radical )( Methyl radical ) Amino group ) Acetic acid

Step (ii) of 1: N-[2- Methoxy radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-N- Methylglycine tert-butyl ester

To a solution of intermediate 35 (0.141 g; 0.42 mmol) and intermediate 1 (0.129 mg; 0.46 mmol) in MeCN (2 mL) was added EDC (0.121 g; 0.63 mmol). The reaction mixture was stirred at ambient temperature for 18 h. The reaction mixture was diluted with pyridine (2 mL) and heated in a microwave at 150 ℃ for 30 minutes. The solvent was removed in vacuo and the residue was dissolved in DCM. The mixture was washed with water and the organic phase was passed through a hydrophobic frit filter plate. The solvent was evaporated in vacuo. The residue was purified by flash chromatography on silica gel, eluting with isohexane/EtOAc (100%/0 to 4:1), to give the title compound. ¹H NMR (CDCl₃, 400MHz) δ 8.63 (1 H, d, J = 1.2 Hz), 8.40 (1 H, dd, J = 8, 1.7 Hz), 8.17-8.15 (2 H, m), 7.55-7.53 (2 H, m), 7.48-7.46 (1 H, m), 7.34-7.16 (4 H, m), 4.11 (1 H, m), 3.74-3.68 (2 H, m), 3.44 (1 H, J = 17.2 Hz), 3.33 (3 H, s), 3.25 (1 H, d, J = 17.2 Hz), 2.45 (3 H, s), 2.07 (3 H, s), 1.47 (9 H, s)。

Step (ii) of 2: 2-((2- Methoxy radical -1-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Ethyl radical )( Methyl radical ) Amino group ) Acetic acid

To the N- [ 2-methoxy-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl group]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]To tert-butyl-N-methylglycinate (0.078 g; 0.13 mmol) was added a solution of HCl in dioxane (4M; 4 mL) and the reaction mixture was stirred at ambient temperature for 18 h and at 80 ℃ for 2 h. The reaction mixture was cooled and the solvent was evaporated in vacuo. The residue was triturated with ether and dried under vacuum. Dissolving the solid in DMSO-d₆To yield the title compound as a colorless gum (0.055 g, 75%).¹H NMR: (DMSO-d₆/D₂O, 400MHz) δ 8.57 (1 H, s), 8.54-8.52 (1 H, d, J = 8 Hz), 8.24 (2 H, d, J = 8 Hz), 7.79 (2 H, d, J = 8 Hz), 7.69 (1 H, d, J = 8 Hz), 7.40-7.37 (2 H, m), 7.33-7.31 (1 H, m), 7.19 (1 H, d, J =7.2 Hz), 4.78 (1H, m), 4.13-4.10 (1H, m), 3.97 (1H, m), 3.87-3.83 (2H, m), 3.36 (3H, s), 2.75 (3H, s), 2.03 (3H, s). LC/MS (method B) 526 (M + H)⁺. HPLC (method C) Rt 9.13 min (purity: 98.2%).

Examples 52: 2-(2- Hydroxy radical -1-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Ethylamino group ) Acetic acid

Step (ii) of 1: N-[2- Hydroxy radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Glycine tert-butyl ester

To a solution of intermediate 36 (0.563 g; 1.27 mmol) and intermediate 1 (0.391 mg; 1.40 mmol) in MeCN (3 mL) was added EDC (0.321 g; 1.67 mmol). The reaction mixture was stirred at ambient temperature for 18 h. The reaction mixture was diluted with pyridine (2 mL) and heated in a microwave at 150 ℃ for 30 minutes. The solvent was removed in vacuo and the residue was dissolved in DCM. The mixture was washed with water and the organic phase was passed through a hydrophobic frit filter plate. The solvent was evaporated in vacuo. The residue was purified by flash chromatography on silica, eluting with isohexane/EtOAc (10:1), to give the title compound. ¹H NMR (CDCl₃, 400 MHz) δ 8.63 (1 H, s), 8.41-8.39 (1 H, dd, J = 7.6, 1.2 Hz), 8.17 (2 H, d, J = 8.4 Hz), 7.54-7.46 (3 H, m), 7.34-7.15 (4 H, m), 3.90-3.88 (1 H, m), 3.74-3.65 (2 H, m), 3.30 (1 H, d, J = 17.2 Hz), 3.13 (1 H, d, J = 17.2 Hz), 2.61 (1 H, br s), 2.07 (3 H, s), 1.46 (9 H, s), 0.91 (9 H, s), 0.06 (3 H, s), 0.05 (3 H, s)。

Step (ii) of 2: 2-(2- Hydroxy radical -1-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Ethylamino group ) Acetic acid

To N- [ 2-hydroxy-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]To tert-butyl glycinate (0.200 g; 0.13 mmol) was added a solution of HCl in dioxane (4M; 4 mL) and the reaction mixture was stirred at 80 ℃ for 1 hour. The reaction mixture was allowed to cool and the solvent was evaporated in vacuo. The residue was purified by preparative HPLC to give the title compound.¹H NMR: (CDCl₃, 400 MHz) δ 8.57 (1 H, d, J = 1.2 Hz), 8.54-8.52 (1 H, m), 8.14 (2 H, d, J = 8 Hz), 7.70 (1 H, d, J = 8 Hz), 7.63 (2 H, d, J = 8.4 Hz), 7.44-7.38 (2 H, m), 7.34-7.30 (1 H, m), 7.21-7.20 (1 H, m), 3.94-3.91 (1 H, m), 3.65-3.56 (1 H, m), 3.52-3.47 (1 H, m), 3.20 (1 H, d, J = 16.8 Hz), 3.06 (1 H, d, J =16.8 Hz), 2.06 (3H, s). LC/MS (method B) 496 (M + H)⁺. HPLC (method D) Rt 3.04 min (purity: 94.9%).

Examples 53: 1-((4-(5-(2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzyl radical )( Methyl radical ) Amino group ) Cyclopentanecarboxylic acid

Step (ii) of 1: (4-(5-(2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Methanol

The title compound was prepared following general procedure 5, starting from intermediate 3 and intermediate 37. It was isolated as a white solid. H NMR (CDCl)₃, 400 MHz) δ8.43 (1 H, s), 8.22-8.15 (3 H, m), 7.53 (2 H, d, J = 8.0 Hz), 7.36-7.24 (4 H, m), 7.13 (1 H, d, J = 7.4 Hz), 4.81 (2 H, d, J = 5.6 Hz), 4.23 (2 H, d, J = 1.8 Hz), 3.33 (3H, s), 2.08 (3H, s), 1.80-1.75 (1H, m). LC/MS (method B) 387 (M + H)⁺. HPLC (method E) Rt =10.9 min (purity: 96.7%).

Step (ii) of 2: 4-(5-(2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzaldehyde

The title compound was prepared according to general procedure 6, starting from (4- (5- (2- (methoxymethyl) -2' -methylbiphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) methanol (750 mg; 1.94 mmol) obtained in step 1. It was isolated as a white solid (732 mg; 98%). H NMR (CDCl)₃, 400 MHz) δ10.12 (1 H, s), 8.45 (1 H, s), 8.40 (2 H, d, J = 8.1 Hz), 8.19 (1 H, dd, J = 7.9, 1.9 Hz), 8.05 (2 H, d, J = 8.2 Hz), 7.37-7.23 (4 H, m), 7.13 (1 H, d, J = 7.4 Hz), 4.27-4.18 (2H, m), 3.34 (3H, s), 2.09 (3H, s). LC/MS (method B) 385 (M + H)⁺. HPLC (method D) Rt =4.71 min (purity: 95.9%).

Step (ii) of 3: 1-((4-(5-(2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzyl radical )( Methyl radical ) Amino group ) Cyclopentanecarboxylic acid

The title compound was prepared according to general procedure 7, starting from 4- (5- (2- (methoxymethyl) -2' -methylbiphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) benzaldehyde (104 mg; 0.27 mmol) obtained in step 2 and using 1-aminocyclopentanoic acid (65 mg; 0.50 mmol). It was isolated as a white solid.¹H NMR (DMSO-d₆, 400 MHz) δ 12.2 (1 H, br s), 8.35 (1 H, d, J = 1.8 Hz), 8.20 (1 H, dd, J = 7.9, 1.9 Hz), 8.16-8.07 (2 H, m), 7.59 (2 H, d, J = 8 Hz), 7.45 (1 H, d, J = 8 Hz), 7.40-7.29 (3 H, m), 7.18 (1 H, d, J = 7.4 Hz), 4.27 (1 H, d, J = 12 Hz), 4.22 (1 H, d, J = 12 Hz), 3.76 (2H, s), 3.28 (3H, s), 2.26-2.21 (5H, m), 2.07 (3H, s), 1.79-1.76 (4H, m), 1.69-1.65 (2H, m). LC/MS (method B): 512 (M + H)⁺. HPLC (method D) Rt 2.81 min (purity: 94.0%).

Examples 54: (2S)-2-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzylamino group ) Butyric acid

Step (ii) of 1: (4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Methanol

The title compound was prepared following general procedure 5, starting from intermediate 1 and intermediate 37. The crude product was recrystallized from EtOAc and gasoline to give the title compound as a white solid.¹H NMR: (CDCl₃, 400MHz) δ 8.63 (1 H, s), 8.40 (1 H, d, J = 8.0 Hz), 8.20 (2 H, d, J = 8.0 Hz), 7.54 (2 H, d, J = 7.9 Hz), 7.47 (1 H, d, J = 7.9 Hz), 7.38-7.21 (3 H, m), 7.16 (1 H, d, J = 7.5 Hz), 4.81 (2 H, d, J = 5.7 Hz), 2.14-1.94 (3H, m). LC/MS (method B): 411 (M + H)⁺. HPLC (method G) Rt = 4.50 min (purity: 97.9%).

Step (ii) of 2: 4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzaldehyde

The title compound was prepared according to general procedure 6, starting from (4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) methanol obtained in step 1. Grinding with gasoline/diethyl ether mixtureThe crude product was triturated to give the title compound as a white solid (5.72 g; 97%). Preparation of H NMR (CDCl)₃, 300 MHz) δ10.13 (1 H, s), 8.64 (1 H, s), 8.43-8.36 (3 H, m), 8.06 (2 H, d, J = 8.0 Hz), 7.49 (1 H, d, J = 8.0 Hz), 7.38-7.21 (3 H, m), 7.15 (1 H, d, J = 7.6 Hz), 2.07 (3H, s). LC/MS (method B) 409 (M + H)⁺. HPLC (method G) Rt = 4.89 min (purity: 95.7%).

Step (ii) of 3: (2S)-2-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzylamino group ) Butyric acid

Sodium cyanoborohydride (34.7 mg; 0.55 mmol) was added to a solution of 4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) benzaldehyde (205 mg; 0.50 mmol) obtained in step 2 and (S) -2-aminobutyric acid (103 mg; 1.00 mmol) in a mixture of methanol (3 mL), DCM (3 mL) and acetic acid (75. mu.l). The mixture was stirred at room temperature overnight, then the solvent was removed in vacuo. The residue was purified by preparative HPLC to give the title compound as a white solid. H NMR (DMSO-d) ₆, 400MHz) δ8.58-8.52 (1 H, m), 8.53-8.48 (1 H, m), 8.12 (2 H, d, J = 8.0 Hz), 7.67 (1 H, d, J = 7.9 Hz), 7.62 (2 H, d, J = 8.0 Hz), 7.41-7.35 (2 H, m), 7.33-7.27 (1 H, m), 7.18 (1 H, d, J = 7.5 Hz), 3.96 (1 H, d, J = 14.1 Hz), 3.78 (1 H, d, J = 14.0 Hz), 3.07-3.00 (1H, m), 2.03 (3H, s), 1.69-1.58 (2H, m), 0.94-0.88 (3H, m). LC/MS (method B) 496 (M + H)⁺. HPLC (method D) Rt = 3.16 min (purity: 99.6%).

By the same protocol as example 54, using amino acids appropriate for each example, the examples shown in the following table were prepared:

examples 69: (2S)-2-( Methyl radical (4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzyl radical ) Amino group ) Butyric acid

Sodium cyanoborohydride (18 mg; 0.28 mmol) was added to a solution of 4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) benzaldehyde (example 54, step 2, 103 mg; 0.25 mmol) and (S) -2-aminobutyric acid (52 mg; 0.50 mmol) in a mixture of methanol (3 mL), DCM (3 mL) and acetic acid (38. mu.l). The mixture was stirred at room temperature overnight and filtered through a glass frit filter plate under positive pressure. To the filtrate was added formaldehyde (37% aqueous; 204 mg, 2.51 mmol) followed by AcOH until the pH was in the range of 3-4 (240 mL). To the resulting mixture was added sodium cyanoborohydride (79 mg, 1.26 mmol), the mixture was stirred for 16 hours, the solvent was removed in vacuo, and the residue was purified by reverse phase HPLC to give a white solid as the title compound. H NMR (DMSO-d) ₆, 400 MHz) δ8.57-8.53 (1 H, m), 8.52-8.48 (1 H, m), 8.10 (2 H, d, J = 8.0 Hz), 7.67 (1 H, d, J = 7.9 Hz), 7.57 (2 H, d, J = 8.0 Hz), 7.40-7.35 (2 H, m), 7.32-7.27 (1 H, m), 7.18 (1 H, d, J = 7.5 Hz), 3.86 (1 H, d, J = 14.3 Hz), 3.72 (1 H, d, J = 14.3 Hz), 3.15 (1 H, t, J = 7.4 Hz), 2.24 (3 H, s), 2.03 (3 H, s), 1.76-1.60 (2 H, m), 0.95 (3 H, t, J = 7.3 Hz). LC/MS (method B) 510 (M + H)⁺. HPLC (method D) Rt 3.39 min (purity: 99.8%).

By the same protocol as example 69, using amino acids suitable for each example, the examples shown in the following table were prepared:

examples 76: 3- Methoxy radical -2-{3-[5-(2'- Methyl radical -2- Trifluoromethyl radical - Biphenyl -4- Base of )-[1,2,4] Oxadiazoles as fungicides -3- Base of ]- Benzylamino group }- Propionic acid

Step (ii) of 1: (3-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Methanol

The title compound was prepared following general procedure 5 starting from intermediate 1 and intermediate 38 and isolated as a white solid. Preparation of H NMR (CDCl)₃, 400 MHz) δ8.65-8.58 (2 H, m), 8.40 (1 H, d, J = 7.9 Hz), 8.22 (1 H, s), 8.13 (1 H, d, J = 7.3 Hz), 7.60-7.50 (2 H, m), 7.47 (1 H, d, J = 7.9 Hz), 7.37-7.20 (2 H, m), 7.15 (1 H, d, J = 7.5 Hz), 4.83 (2 H, d, J = 4.2 Hz), 2.07 (3H, s), 1.93-1.81 (1H, m). LC/MS (method B): 411 (M + H)⁺. HPLC (method G) Rt 4.54 min (purity: 96.8%).

Step (ii) of 2: 3-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzaldehyde

The title compound was prepared according to general procedure 6 starting from (3- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) methanol (452 mg; 1.10 mmol) and isolated as a white solid (392 mg; 87%). Preparation of H NMR (CDCl)₃, 400 MHz) δ10.16 (1 H, s), 8.72-8.71 (1 H, m), 8.68-8.62 (1 H, m), 8.48 (1 H, dt, J = 7.7, 1.4 Hz), 8.43-8.40 (1 H, m), 8.09 (1 H, dt, J = 7.7, 1.4 Hz), 7.73 (1 H, t, J = 7.7 Hz), 7.49 (1 H, d, J = 7.9 Hz), 7.35-7.24 (3 H, m), 7.16 (1 H, d, J = 7.5 Hz), 2.07 (3H, s). LC/MS (method B) 409 (M + H)⁺. HPLC (method G) Rt 4.68 min (purity: 97.4%).

Step (ii) of 3: 3- Methoxy radical -2-{3-[5-(2'- Methyl radical -2- Trifluoromethyl radical - Biphenyl -4- Base of )-[1,2,4] Oxadiazoles as fungicides -3- Base of ]- Benzylamino group }- Propionic acid

The title compound was prepared following the same protocol as described for example 54, step 3, starting from 4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) benzaldehyde and O-methylserine. It was isolated as an off-white solid. LC/MS (method B): 512 (M + H)⁺. HPLC (method G) Rt 6.75 min (purity: 97.3%).

By the same protocol as example 76, using amino acids or amino esters suitable for each example, the examples shown in the following table were prepared:

examples 83: 2- Methyl radical -2-( Methyl radical -{3-[5-(2'- Methyl radical -2- Trifluoromethyl radical - Biphenyl -4- Base of )-[1,2,4] Oxadiazoles as fungicides -3- Base of ]- Benzyl radical }- Amino group )- Propionic acid

The title example was prepared following the same protocol as described for example 69, starting from 4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) benzaldehyde and 2-methylalanine. It was isolated as an off-white solid. LC/MS (method B): 508 (M-H)^-。510 (M+H)⁺. HPLC (method D) Rt3.35 min (purity: 99.8%).

By the same protocol as example 83, using amino acids or amino esters suitable for each example, the examples shown in the following table were prepared:

examples 87: (R)-2-( Ethyl radical -{3-[5-(2'- Methyl radical -2- Trifluoromethyl radical - Biphenyl -4- Base of )-[1,2,4] Oxadiazoles as fungicides -3- Base of ]- Benzyl radical }- Amino group )-3- Hydroxy radical - Propionic acid

The title compound was prepared following the same procedure as described for example 69, starting from 4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) benzaldehyde, acetaldehyde and D-serine. It was isolated as an off-white solid. LC/MS (method B) 524 (M-H)^-。526 (M+H)⁺. HPLC (method D) Rt 3.35 min (purity: 99.0%).

Examples 88: 2-( Ethyl radical -{3-[5-(2'- Methyl radical -2- Trifluoromethyl radical - Biphenyl -4- Base of )-[1,2,4] Oxadiazoles as fungicides -3- Base of ]- Benzyl radical }- Amino group )-2- Methyl radical - Propionic acid

The title compound was prepared according to the same protocol as described for example 87, substituting 2-methylalanine for D-serine. It was isolated as a pale yellow oil. LC/MS (method B): 522 (M-H)^-。524 (M+H)⁺. HPLC (method D) Rt 3.49 min (purity: 99.1%).

Examples 89: (2S)-2-( Ethyl radical (4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Benzyl radical ) Amino group ) Propionic acid

The title compound was prepared according to the same scheme as described for example 69 but using acetaldehyde (134 mg; 3.06 mmol) and (S) -2-aminopropionic acid (55 mg; 0.62 mmol). It was isolated as a white solid. H NMR (DMSO-d)₆, 400 MHz) δ 8.56 (1 H, s), 8.52 (1 H, d, J = 8.0 Hz), 8.11 (2 H, d, J = 7.9 Hz), 7.68 (1 H, d, J = 7.9 Hz), 7.63 (2 H, d, J = 8.0 Hz), 7.44-7.37 (2 H, m), 7.34-7.29 (1 H, m), 7.20 (1 H, d, J = 7.5 Hz), 3.92 (1 H, d, J = 15.0 Hz), 3.79 (1 H, d, J = 15.1 Hz), 3.55-3.44 (1 H, m), 2.72-2.57 (2 H, m), 2.05 (3 H, s), 1.26 (3 H, d, J = 7.0 Hz), 1.06-0.97 (3 H, t, J = 7.0 Hz). LC/MS (method B) 510 (M + H)⁺. HPLC (method D) Rt 3.38 min (purity: 98.6%).

Examples 90: (R)-2-( Ethyl radical -{4-[5-(2'- Methyl radical -2- Trifluoromethyl radical - Biphenyl -4- Base of )-[1,2,4] Oxadiazoles as fungicides -3- Base of ]- Benzyl radical }- Amino group )- Propionic acid

The title compound was prepared according to the same protocol as described for example 89, substituting D-alanine for (S) -2-aminopropionic acid. It was isolated as a colorless oil. LC/MS (method B): 508 (M-H)^-。510 (M+H)⁺. HPLC (method D) Rt = 3.39 min (purity: 99.08%).

Examples 91: (2S)-3- Methyl radical -2-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenethylamino group ) Butyric acid

Step (ii) of 1: (4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Ethanol

The title compound was prepared following general procedure 5, starting from intermediate 1 and intermediate 39. It was isolated as a white solid. Preparation of H NMR (CDCl)₃, 400 MHz) δ8.63 (1 H, s), 8.40 (1 H, dd, J = 8.0, 1.7 Hz), 8.15 (2 H, d, J = 8.1 Hz), 7.47 (1 H, d, J = 8.0 Hz), 7.41 (2 H, d, J = 8.0 Hz), 7.39-7.22 (3 H, m), 7.15 (1 H, d, J = 7.6 Hz), 3.94 (2 H, s), 2.97 (2 H, t, J = 6.5 Hz), 2.06 (3H, s), 1.44 (1H, s). LC/MS (method B) 425 (M + H)⁺. HPLC (method G) Rt 4.31 min (purity: 99.3%).

Step (ii) of 2: 3- Methyl radical -2-(4-(5-(2 ’ - Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenethylamino group ) Butyric acid -(2S)- Tert-butyl ester

To (4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) ethanol (84 mg; 0.2 mmol) in DCM (2 mL) at 0 ℃ was added DIEA (71 μ L;0.4 mmol) and methanesulfonyl chloride (17 μ L; 0.22 mmol). The resulting mixture was stirred at 0 ℃ for 10 minutes, warmed to room temperature, and stirred for 2 hours. The mixture was diluted with DCM (20 mL) and saturated NaHCO was added ₃An aqueous solution. The aqueous layer was extracted with DCM (3X20 mL) and dried (MgSO)₄) The combined organic fractions were filtered and the solvent was removed in vacuo. All crude residues were used in the next step without further purification. It was dissolved in dioxane (2 mL). Potassium carbonate (165 mg, 1.2 mmol) and 2-amino-3-methylbutyric acid- (S) -tert-butyl ester (104 mg; 0.60 mmol) were added. The mixture was heated at 130 ℃ for 72 h and diluted with DCM (5 mL) and water (5 mL). The aqueous layer was extracted with DCM (3X20 mL) and dried (MgSO)₄) The combined organic fractions were filtered and the solvent was removed in vacuo. The residue was purified by flash chromatography, eluting with petrol containing increasing amounts of EtOAc, to give the title compound as a colourless gum (86 mg; 72%). Preparation of H NMR (CDCl)₃, 400 MHz) δ8.63 (1 H, s), 8.39 (1 H, dd, J = 8.0, 1.7 Hz), 8.12 (2 H, d, J = 8.1 Hz), 7.47 (1 H, d, J = 8.0 Hz), 7.43-7.21 (5 H, m), 7.16 (1 H, d, J = 7.6 Hz), 2.99-2.68 (5 H, m), 2.07 (3 H, s), 1.94-1.79 (1 H, m), 1.46 (9 H, s), 0.94 (6 H, dd, J = 6.8, 3.7 Hz). LC/MS (method B): 580 (M + H)⁺. HPLC (method D) Rt 3.7 min (purity: 94.9%).

Step (ii) of 3: (2S)-3- Methyl radical -2-(4-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenethylamino group ) Butyric acid

To 3-methyl-2- (4- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenethylamino) butanoic acid- (2S) -tert-butyl ester (86 mg; 0.15 mmol) was added 4M HCl in dioxane and the mixture was heated in a test tube at 70 ℃ for 3 hours. Then is at The solvent was removed in vacuo and the residue was purified by preparative HPLC to give the title compound as a white solid. H NMR (DMSO-d)₆, 400 MHz) δ8.56 (1 H, s), 8.50 (1 H, d, J = 8.1 Hz), 8.08 (2 H, d, J = 7.8 Hz), 7.65 (1 H, d, J = 8.1 Hz), 7.51 (2 H, d, J = 8.0 Hz), 7.40-7.38 (2 H, m), 7.31 (1 H, td, J = 7.0, 2.3 Hz), 7.18 (1 H, d, J = 7.6 Hz), 3.09-2.80 (5 H, m), 2.16 (3 H, s), 1.99-1.90 (1 H, m), 0.96 (6 H, dd, J = 6.8, 2.1 Hz). LC/MS (method B) 524 (M + H)⁺. HPLC (method H) Rt 9.20 min (purity: 97.7%).

By the same protocol as example 91, using amino acids or amino esters suitable for each example, the examples shown in the following table were prepared:

examples 94: (2S)-2-(3-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenethylamino group ) Propionic acid

Step (ii) of 1: (3-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenyl radical ) Ethanol

The title compound was prepared according to general procedure 5 starting from intermediate 1 and intermediate 40 and isolated as a white solid (1.03 g, 80%). Preparation of H NMR (CDCl)₃400 MHz) δ 8.66 (s, 1H), 8.42 (dd, J = 8.0, 1.7 Hz, 1H), 8.11-8.08 (m, 2H), 7.54-7.21 (m, 6H), 7.17 (d, J = 7.7 Hz, 1H), 3.98 (dd, J = 12.1, 6.0 Hz, 2H), 3.02 (t, J = 6.5 Hz, 2H), 2.08 (s, 3H), 1.50-1.43 (m, 1H). LC/MS (method B) 425 (M + H)⁺. HPLC (method G) Rt 4.30 min (purity: 95.8%).

Step (ii) of 2: (2S)-2-(3-(5-(2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of )-1,2,4- Oxadiazoles as fungicides -3- Base of ) Phenethylamino group ) Propionic acid

The title compound was prepared according to the protocol used for example 91, steps 2 and 3 but starting from (3- (5- (2' -methyl-2- (trifluoromethyl) biphenyl-4-yl) -1,2, 4-oxadiazol-3-yl) phenyl) ethanol (84 mg, 0.2 mmol) and- (S) -tert-butyl 2-aminopropionate (87 mg, 0.6 mmol). It was isolated as a white solid (133 mg, 96%). H NMR (DMSO-d) ₆400 MHz) δ 8.57 (s, 1H), 8.53 (d, J = 8.1 Hz, 1H), 8.07-8.03 (m, 2H), 7.69 (d, J = 8.0 Hz, 1H), 7.65-7.53 (m, 2H), 7.44-7.37 (m, 2H), 7.33 (td, J = 4.0, 2.1 Hz, 1H), 7.20 (d, J = 7.6 Hz, 1H), 3.31 (q, J = 7.2 Hz, 2H), 3.18-3.02 (m, 3H), 2.06 (s, 3H), 1.33 (d, J = 7.0 Hz, 3H). LC/MS (method B) 496 (M + H)⁺. HPLC (method G) Rt 3.24 min (purity: 99.1%).

By the same protocol as example 94, using amino acids or amino esters suitable for each example, the examples shown in the following table were prepared:

examples 97: N-[(1R)-1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-N- Methyl glycine , Hydrochloride salt

Preparation of N- [ (1R) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 24 and intermediate 3 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-N-methylglycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆, 400 MHz) δ 8.31(s, 1H), 8.20-8.14 (m, 3H) 7.81 (d, J = 8.2 Hz, 2H), 7.42 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 3.8 Hz, 2H), 7.29 (q, J = 5.5 Hz, 2H), 7.13 (d, J = 7.0 Hz, 1H), 4.72 (d, J = 6.4 Hz, 1H), 4.17 (dd, J = 12.7 Hz, 2H ), 4.00-3.92 (m, 2H), 3.24 (s, 3H), 2.75 (s, 3H), 2.02 (s, 3H ), 1.68 (d, J = 6.7 Hz, 3H). LC/MS (method A): 472.3 (M + H)⁺, 526.3 (M-H)^-. HPLC (method A) Rt 4.62 min (purity: 97.8%).

Examples 98: N-[(1S)-1-(4-{5-[2-( Methoxymethyl group )-2'- Methyl biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-N- Methyl glycine , Hydrochloride salt

Preparation of N- [ (1S) -1- (4- {5- [2- (methoxymethyl) -2' -methylbiphenyl-4-yl) starting from intermediate 23 and intermediate 3 according to general procedure 3]-1,2, 4-oxadiazol-3-yl } phenyl) ethyl]-N-methylglycine tert-butyl ester. It was hydrolyzed according to general procedure 8 to give the title compound as a white powder.¹H NMR: (DMSO-d₆, 400 MHz) δ 8.31(s, 1H), 8.21-8.15 (m, 3H), 7.81 (d, J = 8.2 Hz, 2H), 7.42 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 4.1 Hz, 2H), 7.30-26 (m, 1H), 7.13 (d, J = 7.2Hz, 1H), 4.72 (d, J = 5.9 Hz, 1H), 4.23-4.14 (dd, J = 12.7 Hz, 2H), 3.97-3.96 (m, 2H), 3.24 (s, 3H), 2.75 (s, 3H), 2.02(s, 3H), 1.68 (d, J = 6.8 Hz, 3H). LC/MS (method A): 472.3 (M + H)⁺, 526.3 (M-H)^-. HPLC (method A) Rt 4.64 min (purity: 98.8%).

Examples 99 And examples 100: N-[(1S)-2- Hydroxy radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt and N-[(1R)-2- hydroxy radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

Step (ii) of 1: N-[2-{[ Tert-butyl radical ( Dimethyl group ) Silyl radical ] Oxy radical }-1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine tert-butyl ester

To a solution of intermediate 46 (2 g, 0.0045 mol) in DMF (30 ml) at room temperature was added intermediate 1 (1.54 g, 0.0054 mol), triethylamine (2.5 ml, 0.018 mol) and T₃P (3.57 g, 0.0112 mol). The reaction mixture was heated at 70 ℃ for 15 h. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in ethyl acetate, water and 10% NaHCO₃Washing with Na solution₂SO₄Dried and evaporated to give (1.2 g, 38%) the title compound as a brown oil. The crude material was used in the next step without purification. LC/MS (method A) 682.3 (M + H) ⁺. HPLC (method A)Rt 7.1 min (purity: 70.3%).

Step (ii) of 2: N-[(1S)-2- Hydroxy radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt and N-[(1R)-2- hydroxy radical -1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ]-D- Alanine , Hydrochloride salt

To a solution of N- [2- { [ tert-butyl (dimethyl) silyl ] oxy } -1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine tert-butyl ester (1.2 g, 0.0017 mol) in THF (25 ml) was added TBAF (50 mg, 0.1 equiv.) at room temperature. The reaction mixture was stirred at room temperature for 30 min, quenched with water, extracted with ethyl acetate, and concentrated under reduced pressure. The crude product was purified by column chromatography using chloroform and methanol (90:10) as eluent to give N- [ 2-hydroxy-1- (4- {5- [2' -methyl-2- (trifluoromethyl) biphenyl-4-yl ] -1,2, 4-oxadiazol-3-yl } phenyl) ethyl ] -D-alanine tert-butyl ester as a brown oil as a mixture of 2 diastereomers (0.7 g, 77%). The 2 diastereomers were separated by chromatography (SFC on Chiralpak ADH, 20% cosolvent: 0.5% DEA in ethanol, total flow: 3 ml/min), the Rt of the first eluting isomer being at 4.06 min and the Rt of the subsequent eluting isomer being at 5.25 min.

Each diastereomer was dissolved in 4M HCl (in dioxane, 4 ml). The resulting mixture was stirred at room temperature for 10 h, then heated at 80 ℃ for 2 h. Then concentrated under reduced pressure. The crude product was purified by prep-HPLC using water/acetonitrile as eluent to give the title compound as an off-white solid.

Diastereomer eluting first, after hydrolysis: 50 mg, 43%

¹H NMR: (DMSO-d₆, 400 MHz) δ 8.51 (s, 1H), 8.50-8.45 (d, J = 7.9 Hz, 1H), 8.12-8.08 (d, J = 8.2 Hz, 2H), 7.66-7.58 (m, 3H), 7.38-7.33 (m, 2H), 7.30-7.24 (m, 1H), 7.17-7.13 (d, J = 7.9 Hz, 1H), 4.92-4.88 (m, 1H), 3.28-3.24 (m, 1H), 3.03-2.97 (m, 1H), 2.90-2.80 (m, 1H), 2.00 (s, 3H), 1.30-1.26 (d, J= 7.2 Hz, 3H). LC/MS (method A) 512.3 (M + H)⁺. HPLC (method A) Rt 4.83 min (purity: 99.2%).

The diastereomer subsequently eluted, after hydrolysis: 50 mg, 43%

¹H NMR: (DMSO-d₆, 400 MHz) δ 8.51 (s, 1H), 8.50-8.45 (d, J = 7.9 Hz, 1H), 8.12-8.08 (d, J = 8.2 Hz, 2H), 7.66-7.58 (m, 3H), 7.38-7.33 (m, 2H), 7.30-7.24 (m, 1H), 7.17-7.13 (d, J = 7.9 Hz, 1H), 4.97-4.94 (m, 1H), 3.37-3.33 (m, 1H), 3.08-3.03 (m, 1H), 2.95-2.80 (m, 1H), 2.01 (s, 3H), 1.31-1.27 (d, J= 7.2 Hz, 3H). LC/MS (method A) 512.3 (M + H)⁺. HPLC (method A) Rt 4.90 min (purity: 99.4%).

Examples 101: 1-[ Methyl radical (4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical ) Nitroso claw radical ] Cyclopentanecarboxylic acid

The title compound was prepared according to general procedure 12 starting from example 53(95.7 mg; 0.18 mmol). After evaporation of the solvent, the crude mixture was triturated in DMSO/MeCN 1:1 mixture (2 mL), filtered, and washed with MeCN to give the title compound as a white solid (69.50 mg; 70.5%).¹H NMR: (CDCl₃, 300 MHz) δ 8.65 (br d, J = 1.4 Hz, 1H), 8.43 (br dd, J = 1.4, 8.0 Hz, 1H), 8.35-8.28 (m, 2H), 7.78-7.70 (m, 2H), 7.51 (d, J = 8.0 Hz, 1H), 7.41-7.23 (m, 3H), 7.17 (br d, J = 7.6 Hz, 1H), 4.97 (br d, J = 12.6 Hz, 1H), 4.48 (br d, J= 12.6 Hz, 1H), 3.06 (s, 3H), 2.96-2.70 (m, 1H), 2.70-2.32 (m, 2H), 2.20-1.66 (m, 5H), 2.08 (s, 3H). LC/MS (method A) 552.5 (M + H) ⁺。550.5 (M+H)^-. HPLC (method A) Rt 5.15 min (purity: 98.4%).

Examples 102: 2-[(2- Fluorine -4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Benzyl radical )( Methyl radical ) Nitroso claw radical ] Propionic acid

Example 50 (100 mg; 0.18 mmol) was dissolved in EtOAc and washed with NaHCO₃Washing with saturated solution. Glacial acetic acid (1 mL) was added and the resulting solution was washed with water and brine. Over MgSO₄Dried, filtered and evaporated. Then, starting from the precursor of example 50 (92.4 mg; 0.18 mmol), the general procedure 12 is followed. After evaporation of the solvent, the crude mixture was dissolved in a 1:1 mixture of DMSO/MeCN (1 mL) and purified by MD-autoprep. The title compound was isolated as a white solid as a 47:53 diastereomeric mixture (LC/MS method a).¹H NMR: (CDCl₃, 300 MHz) δ 8.64 (br d, J = 1.5 Hz, 1H), 8.42 (br dd, J= 1.5, 8.0 Hz, 1H), 8.18-8.12 (m, 1H), 8.10-8.03 (m, 1H), 7.99-7.84 (m, 1H), 7.51 (d, J = 8.1 Hz, 1H), 7.41-7.23 (m, 3H), 7.16 (br d, J = 7.4 Hz, 1H), 4.87 and 4.81 (2 br s, 2H), 4.47-4.31 and 4.24-4.07 (2 m, 1H), 3.28 and 3.19 (2 s, 3H), 2.08 (s, 3H), 1.92 and 1.80 (2 d, J = 6.9 Hz, 3H). LC/MS (method A): 2 Rt for diastereomer at 1.87 min (47%) and 1.94 min (53%): 530.4 (M + H)⁺. HPLC (method A) Rt 4.84 min (purity: 94.1%).

Examples 103: { Methyl radical [1-(4-{5-[2'- Methyl radical -2-( Trifluoromethyl radical ) Biphenyl -4- Base of ]-1,2,4- Oxadiazoles as fungicides -3- Base of } Phenyl radical ) Ethyl radical ] Nitroso claw radical } Acetic acid

Example 40 (50 mg; 0.09 mmol) was dissolved in EtOAc and washed with NaHCO₃Washing with saturated solution. Glacial acetic acid (1 mL) was added and the resulting solution was washed with water and brine. It is passed over MgSO₄Dried, filtered and evaporated. Then, starting from the precursor of example 40 (44.6 mg; 0.09 mmol), the general procedure 12 is followed. After evaporation of the solvent, the crude mixture was dissolved in a 1:1 mixture of DMSO/MeCN (1 mL) and purified by MD-autoprep. The title compound was isolated as a white solid as a 22:78 diastereomeric mixture (LC/MS method a).¹H NMR: (CDCl₃300 MHz) δ 8.68-8.61 (m, 1H), 8.50-8.08 (m, 3H), 7.84-7.56 (m, 2H), 7.55-7.45 (m, 1H), 7.42-7.22 (m, 3H), 7.17 (br d, J = 6.7 Hz, 1H), 5.47-5.27 (m, 1H), 4.80 (br d, J = 14.2 Hz, 1H), 4.62 (br d, J = 14.2 Hz, 1H), 3.90 and 3.48 (2 s, 3H), 2.08 (s, 3H), 2.02-1.88 (m, 3H). LC/MS (method A): 2 Rt for diastereomer at 1.87 min (22%) and 1.94 min (78%): 512.5 (M + H)⁺. HPLC (method A) Rt 5.15 min (purity: 98.4%).

Examples 104: In vitro assay

Membrane preparation: Expression of S1P₁Or S1P₃CHO cell of (e), to prepare a membrane for ligand binding studies. The cells were suspended in 50 mM TRIS (pH 7.4), 2 mM EDTA, 250 mM sucrose (buffer A) and 1 Xcomplete protease inhibitor mixture (Roche) and disrupted by N2 under reduced pressure using a cell disruption bomb (Parr Instrument) at 4 ℃. After centrifugation at 1000 RPM for 10 min at 4 deg.C, the supernatant was diluted (2X) in buffer A and centrifuged again for 75 min at 19000 RPM at 4 deg.C. The pellet was then suspended in 10 mM HEPES (pH 7.4), 1 mM EDTA, 250 mM sucrose (buffer B) and 1 Xcomplete EDTA-free protease inhibitor cocktail and homogenized using a potter. The membranes were snap frozen in liquid nitrogen and stored at-80 ℃.

Receptor binding assays: will [33P ]]Sphingosine 1-phosphate (3000 Ci/mmol; American radio laboratory Chemicals, Inc.) was added to the test compound in 20% DMSO for competition. Membranes and WGA SPA beads (GE Healthcare) were added to a final volume of 100 μ l (in 96-well plates) or 50 μ l (in 384-well plates) at the determined concentrations: 30 pM or 15 pM [33P ]]Sphingosine 1-phosphate (for S1P, respectively)₁Or S1P₃)、50 mM HEPES（pH 7.5）、5 mM MgCl₂100 mM NaCl, 0.4% fatty acid free BSA, 1-5 μ g/well of protein (in 96-well plates) or 0.6-1 μ g/well of protein (in 384-well plates), and 100 μ g/well of WGA SPA beads (in 96-well plates) or 75 μ g/well of WGA SPA beads (in 384-well plates). Binding was performed on a shaker at room temperature for 60 min and the bound radioactivity was measured on a PerkinElmer 1450 MicroBeta counter. Triplicate samples were averaged and normalized to percent inhibition relative to total binding (DMSO only in wells) and non-specific binding (1000-fold excess unlabeled S1P). Binding data was analyzed using GraphPad Prism program or Genedata software.

Cell function assay: in that 384- Human cell line in pore form (U2OS) Sphingosine of (A) -1- Phosphate receptor ₁1 (S1P) Internalization of (analysis using cell imaging)

Jo, E.; Sanna, M. G.; Gonzalez-Cabrera, P. J.; Thangada, S.; Tigyi, S.; Osborne, D. A.; Hla, T.; Parrill, A. L.; Rosen, H. Chem. Biol. 2005, 12, 703

S1P from BioImage was used₁U2OS cells (C039A) (human epithelial cell line (human bone osteosarcoma epithelial cells)), S1P in 384 well plates (Corning ® 384 black with clear bottom 3712)₁And (4) internalization determination. These cells express human S1P fused to Green fluorescent protein (EGFP)₁A receptor. Standard CMV promoter (Cytomegalovirus promoter) control S1P₁Expression of EGFP by adding GenetiaThe elements maintain continuous expression.

S1P₁Receptor desensitization induces membrane-localized S1P₁Internalization of EGFP fusion proteins into endosomes, which can be monitored by cellular imaging analysis.

Cells were plated overnight in low serum medium (Dulbecco's modified eagle's medium (DMEM) containing Glutamax-1 and high glucose, 1% penicillin/streptomycin, 1% Fetal Calf Serum (FCS), 0.5 mg/ml geneticin).

The next day, at 37 deg.C/5% CO₂S1P was incubated in 20 μ l serum-free medium (DMEM with Glutamax-1 and high glucose, 0.1% fatty acid-free Bovine Serum Albumin (BSA) 10 mM, N '-2-hydroxyethylpiperazine-N' -2 ethanesulfonic acid (HEPES) 1M) ₁U2OS cells for 2 hours.

Cells (total volume 24 μ l) were then treated with 4 μ l compound/agonist (6x/3% DMSO) and CO at 37 ℃/5%₂The plates were incubated for 1 hour.

Fixing S1P with 25 mul 8% paraformaldehyde₁U2OS cells and stained with Hoechst 33345 dye (1:1000) for 20 min.

They were then washed 3 times with Phosphate Buffered Saline (PBS) and the plates were sealed.

On Cellomics, the "number of blobs per object" was calculated (object corresponds to nucleus, blob corresponds to S1P₁EGFP receptor), the measurement receptor S1P₁Internalization of EGFP. The internalized data were observed with the aid of vHCS View and analyzed using Genedata software.

The compounds of formula (I) can be used as immunomodulators, as they act as₁P₁The activity of an effective agonist of the receptor (as measured in the above assay) is demonstrated. A compound of the formulae (I) and (la) to S₁P₁Ki of less than 0.1. mu.M. Preferred compounds of formula (I) exhibit an affinity for S₁P₁Ki of less than 0.01. mu.M for the receptor. More preferred is the conversion of formula (I)The compound shows a structure of a pair S₁P₁Ki of less than 0.001. mu.M. The compounds of formula (I) exhibit an affinity for S₁P₁Superiority of the receptor over S₁P₃Selectivity of the receptor, just as by the pair S₁P₁Ki and pair S of receptors₁P₃The ratio of Ki for the receptors (as assessed in the binding assay described above) was determined. Ki S ₁P₁And Ki S₁P₃The ratio of (d) is greater than 20, preferably greater than 50, more preferably greater than 100 and even more preferably greater than 1000.

The "potency" or "activity" of a compound is determined by the Ki value, which is evaluated in one of the above binding assays (96 or 384 well plates). The lowest Ki value characterizes the most potent or active compound according to the invention.

The following results have been obtained:

examples 105 : evaluation of S1P Animal models of in vivo efficacy of agonists

S1P Agonist-induced lymphopenia mouse model

Female C57BL/6 mice (Elevage Janvier) (8 weeks old) received S1P agonist by oral route. Blood samples of heparinized (100 IU/kg, intraperitoneal) mice were drawn under isoflurane anesthesia by intracardiac or retroorbital puncture 2-120 hours after drug treatment. White blood cells (lymphocytes and neutrophils) were counted using a Beckman/Coulter counter. The quality of the blood sample was assessed by counting red blood cells and platelets.

MOG Induced experimental autoimmune encephalomyelitis (EAE) Mouse model

EAE was induced in 9-week-old female mice (C57BL/6, Elevage Janvier) by immunization against MOG. The mice received pertussis toxin (Alexis, 300 ng/mouse in 200 μ l PBS) by intraperitoneal route and 100 μ l of emulsion containing MOG35-55 peptide (NeoMPS, 200 μ g/mouse), Mycobacterium tuberculosis (0.25 mg/mouse) in complete Freund's adjuvant (DIFCO) by subcutaneous injection into the back. After 2 days, pertussis toxin (Alexis, 300 ng/mouse in 200 μ l PBS) was re-injected by intraperitoneal route. Mice were weighed daily after EAE induction and neurological lesions quantified by assessing paralysis (tail, hind limb and forelimb), incontinence and death using a 15-point clinical scale.

Pharmacokinetic data:

the pharmacokinetic properties of the compound 2- ({ 2-fluoro-4- [5- (2' -methyl-2-trifluoromethyl-biphenyl-4-yl) - [1,2,4] oxadiazol-3-yl ] -benzyl } -methyl-amino) -2-methyl-propionic acid of example 31 are as follows:

lymphocyte depletion in mice at 48 h%	67+/-6
		PK parameters from lymphopenia (mice)	PK-PD (30mpk)
Plasma AUC ∞ (h ng/ml)	74978
		Cmax (ng/ml)	7423
Tmax(h)	2
		T1/2 (h)	6.3
Cl/F (L/kg/h)	0.4
		Brain/plasma ratio: AUCZ (h xing ng/ml)	3.5
Lymph node/plasma ratio: 24h/48h	2.5/2.7

Clinical scoring

-1-tail

Score = 0 normal mice kept the tail upright while exercising.

Score = 1 there was a tendency to fall if the tail end was loose.

Score = 2 if the tail is completely relaxed and dragged on a table.

-2-hind limb

Score = 0 normal mice had a vigorous gait, without dragging their paw.

Score = 1 one of the following tests was positive:

a-flick test: while holding the tail between the thumb and forefinger, the animal's back was flicked and the time it took to erect himself was observed. Healthy mice will immediately turn themselves over. The delay suggests hind limb weakness.

B-the mouse is placed on top of the wire cage and observed as it passes from side to side. We believe that there is partial paralysis if one or both limbs frequently slip between the threadlines.

Score = 2 the first 2 tests were all positive.

Score = 3 one or two hind limbs showed signs of paralysis, but some movement was retained; for example: the animal can grasp and hang for a short time on the underside of the top of the wire cage before falling.

Score = 4 both hind legs were paralyzed and the mice pulled the body while moving.

-3-forelimb:

score = 0 a normal mouse is actively using its forepaws for grasping and walking, and keeping its head upright.

Score = 1 walking was possible but made difficult by the weakness of one or both paws, e.g. the front paw was considered weak when the mouse had difficulty grasping the underside of the top of the wire cage. Another sign of weakness is head droop.

Score = 2 when one forelimb is paralyzed (not able to grip and the mouse turns around the paralyzed limb). At this point, the head has also lost most of the muscle tone.

Score = 3 mice were unable to move and were unable to acquire room temperature and water.

-4-bladder:

score = 0 normal mice had complete control of their bladder.

Score = 1 a mouse was considered incontinent when its lower body was soaked with urine.

-5-death:

score = 15.

By adding all the above categories, the final score for each animal was determined. The maximum score of a live animal is 10.

On day 12 (first sign of paralysis), mice in the experimental group (n = 10) were graded according to clinical score and weight loss. Semi-therapeutic treatment was started on day 14.

Examples 106 : preparation of pharmaceutical formulations

Preparation 1 - And (3) tablet preparation:the compound of formula (I) is mixed with a dry gelatin binder in a dry powder ratio of about 1:2 by weight. A small amount of magnesium stearate was added as a lubricant. The mixture is formulated in a tablet press as 240-270 mg tablets (80-90 mg of active compound according to the invention per tablet).

Preparation 2 - And (3) capsule preparation:the compound of formula (I) is mixed with a starch diluent in a dry powder ratio of about 1:1 by weight. The mixture is filled into 250 mg capsules (125 mg of active compound according to the invention per capsule).

Preparation 3 - Liquid:the compound of formula (I) (1250 mg), sucrose (1.75 g) and xanthan gum (4 mg) were blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously prepared solution of microcrystalline cellulose and sodium carboxymethylcellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavours and colours were diluted with water and added with stirring. Sufficient water was then added to generate a total volume of 5 mL.

Preparation 4 - And (3) tablet preparation:the compound of formula (I) is mixed with a dry gelatin binder in a dry powder ratio of about 1:2 by weight. A small amount of magnesium stearate was added as a lubricant. The mixture was formulated into 450-900 mg tablets (150-300 mg of active compound according to the invention) in a tablet press.

Preparation 5 - Injection preparation: the compound of formula (I) is dissolved in a buffered sterile saline injectable aqueous medium to a concentration of about 5 mg/mL.

Claims (16)

1. Compounds of formula (I) and groups thereinIs oxidized intoIn oxidized form, pharmaceutically acceptable derivatives, solvates, tautomers, salts, enantiomers, diastereomers, including the sameMixtures of all proportions

Wherein

R¹Represents H, halogen, CF₃、OCF₃CN or NO₂、OH、A、OA

X¹、X²、Z¹、Z²Independently of one another, H or A, wherein X¹、X²、Z¹、Z²At least one of which is a,

X¹、X²can be reacted with C₁Together form a 3-8 membered ring,

Z¹、Z²can be reacted with C₂Together form a 4-5 membered ring,

Y¹、Y²h, A or glucuronide independently of each other,

W¹、W²、W³and W⁴Independently of one another represent-CH₂-a group or a single bond,

R^arepresents halogen or a linear or branched alkyl group having 1 to 3 carbon atoms, wherein 1 to 3H atoms may be replaced by halogen,

R^brepresents halogen, straight-chain or branched alkyl having 1 to 3 carbon atoms, in which 1 to 3H atoms may be replaced by halogen, -OCH₃、-OCH₂CH₃Instead of this, the user can,

G¹、G²independently of one another, H, halogen, A,

a represents a linear or branched alkyl group having 1 to 6 carbon atoms, preferably 1 to 3 carbon atoms, in which 1 to 3H atoms may be substituted by halogen, OCH₃And the substitution of OH is carried out,

C₁、C₂is a carbon atom.

2. A compound of formula (I) according to claim 1, wherein X¹And Z¹Are both H, and wherein X²And Z²Represents A, and/or X ²And Z²Are both H, and wherein X¹And Z¹A is represented.

3. A compound of formula (I) according to claim 1, wherein X¹And Z²Are both H, and wherein X²And Z¹Represents A, and/or X²And Z¹Are both H, and wherein X¹And Z²A is represented.

4. A compound of formula (I) according to claims 1-3, wherein groupRepresents one of the following groups:

。

5. a compound of formula (I) according to claim 1, wherein the group R^aSelected from halogen, CH₃、CHF₂、CF₃、CH₂CH₃。

6. A compound of formula (I) according to claim 1, wherein groupAs defined in claim 4, and R^aSelected from halogen, CH₃、CHF₂、CF₃、CH₂CH₃。

7. Compounds of formula (I) according to claims 1 to 6 of the following groups 1 to 96 and pharmaceutically acceptable derivatives, solvates, tautomers, salts and stereoisomers thereof, including mixtures thereof in all ratios:

example numbering Formula (II) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 M1 M2

。

8. A process for the preparation of compounds of the formula (I) and salts thereof according to one or more of claims 1 to 7,

reacting a compound of formula A in the presence of a suitable base or, if T is OH, in the presence of a suitable condensing agent

Wherein G is₁、G₂、R^aAnd R^bHas the meaning given in claim 1 and T is OH or a leaving group, wherein T is OH,

With a compound of the formula B or a diastereomeric and/or enantiomeric mixture thereof

Wherein R is¹、W₁-W₄、C₁、C₂、X₁、X₂、Y₁、Y₂、Z₁And Z₂Having the meaning given in claim 1,

and cyclizing the resulting product;

and optionally converting the base or acid of formula (I) into one of its salts, or any pharmaceutically acceptable derivative, solvate, tautomer, enantiomer, diastereomer thereof, including mixtures thereof in all ratios;

and optionally isolating the resulting product mixture.

9. A pharmaceutical composition comprising at least one compound of formula (I) according to one or more of claims 1 to 7 and/or pharmaceutically acceptable derivatives, tautomers, salts, solvates and stereoisomers thereof, including mixtures thereof in all ratios; and optionally excipients and/or adjuvants.

10. A pharmaceutical composition comprising at least one compound of formula (I) according to one or more of claims 1 to 7 and/or pharmaceutically acceptable derivatives, tautomers, salts, solvates and stereoisomers thereof, including mixtures thereof in all ratios; and at least one other active ingredient.

11. A collection (kit) consisting of the following individual packages:

(a) An effective amount of a compound of formula (I) according to one or more of claims 1 to 6, 9 and 10 and/or pharmaceutically acceptable derivatives, tautomers, salts, solvates and stereoisomers thereof, including mixtures thereof in all ratios,

and

(b) an effective amount of other pharmaceutically active ingredients.

12. A compound according to one or more of claims 1 to 6 and pharmaceutically acceptable derivatives, salts, tautomers, solvates and stereoisomers thereof, including mixtures thereof in all ratios, for the preparation of a medicament for the treatment and/or prevention of sphingosine 1-phosphate related disorders.

13. The compound of claim 12, wherein the sphingosine 1-phosphate- (1) -associated disorder is an autoimmune disorder or disease associated with an overactive immune response.

14. A compound according to one or more of claims 1 to 6 and pharmaceutically acceptable derivatives, salts, tautomers, solvates and stereoisomers thereof, including mixtures thereof in all ratios, for the preparation of a medicament for the treatment and/or prophylaxis of immunoregulatory disorders.

15. The compound of claim 14, wherein the immunoregulatory abnormality is an autoimmune disease or chronic inflammatory disease selected from the group consisting of: systemic lupus erythematosus, chronic rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, Amyotrophic Lateral Sclerosis (ALS), arteriosclerosis, atherosclerosis, scleroderma, autoimmune hepatitis.

16. Formula (I)^M) Or (I)^M2) And pharmaceutically acceptable derivatives, solvates, tautomers, salts, enantiomers, diastereomers thereof, including mixtures thereof in all ratios

Wherein

G³、G⁴、G⁵May represent independently of each other an OH group,

t is 0, 1, 2 or 3,

and G¹、G²、R^a、R^b、R¹、X¹、X²、Y¹、Y²、Z¹、Z²、W¹、W²、W³And W⁴As defined in claim 1.